Polypeptide variants

ABSTRACT

The present invention relates to polypeptide variants and methods for obtaining variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

REFERENCE TO A SEQUENCE LISTING

This application contains a Sequence Listing in computer readable form,which is incorporated herein by reference.

BACKGROUND OF THE INVENTION Field of the Invention

The present invention relates to novel dispersin variants havingimproved properties relative to the parent dispersin, such propertiesmay include one or more of the following; wash performance, stabilitye.g. detergent and/or storage stability. The variants of the inventionare suitable for use in cleaning processes and detergent compositions,such as laundry compositions and dish wash compositions, including handwash and automatic dish wash compositions. The present invention alsorelates to isolated DNA sequences encoding the variants, expressionvectors, host cells, and methods for producing and using the dispersinvariants of the invention.

Description of the Related Art

Enzymes have been used in detergents for decades most commerciallyrelevant are the proteases and amylases effectively removing protein andstarch related soiling, respectively. However, most household carerelated soiling is a complex mixture of various organic matters. Hardsurfaces and fabrics are exposed to skin debris e.g. dead cells, sweat,sebum, pollution, as well as stains derived from microorganisms from theenvironment and from e.g. the body. Complex organic stains, such asbiosoil from human skin e.g. dead skin cells, sweat, sebum in particularunderarm soil, composes different substances such as of protein, starch,grease but also e.g. polysaccharides. Consequently, stain removalrequires different enzyme activity, which vary depending on the specificstain targeted.

To be useful in cleaning processes such as laundry, an enzyme such asdispersin needs to be stable in detergent compositions and compatiblewith standard detergent components such as surfactants, builders,bleaches etc.

The present invention provides such enzymes, which are suitable for usein detergents for e.g. improving cleaning of laundry andwashing/dish-washing machines.

SUMMARY OF THE INVENTION

The present invention relates to dispersin variants, which areparticularly useful in detergents and cleaning processes, such aslaundry and dish wash.

One aspect of the invention relates to a dispersin variant, comprisingan alteration, preferably substitution at one or more position(s)corresponding to position(s): 2, 3, 12, 15, 17, 18, 19, 22, 23, 24, 25,26, 30, 32, 34, 43, 44, 45, 49, 52, 54, 56, 57, 58, 59, 60, 62, 63, 67,68, 71, 72, 74, 77, 79, 80, 81, 82, 90, 99, 100, 103, 104, 105, 106,107, 110, 111, 113, 114, 116, 117, 118, 119, 120, 122, 123, 124, 125,126, 127, 128, 131, 135, 138, 139, 140, 142, 145, 147, 148, 149, 150,151, 152, 163, 164, 167, 168, 170, 171, 173, 174, 175, 177, 178, 179,181, 185, 186, 187, 188, 189, 199, 200, 203, 204, 205, 207, 208, 210,215, 217, 218, 221, 222, 224, 225, 227, 230, 232, 233, 234, 235, 237,244, 249, 251, 252, 253, 254, 256, 260, 261, 262, 263, 264, 265, 267,268, 270, 271, 272, 273, 274, 276, 278, 279, 280, 281, 282, 283, 284,287, 288, 290, 291, 296, 300, 301, 303, 304, 305, 306, 308, 309, 312,314, 315, 319, 321 or 323 of the polypeptide shown in SEQ ID NO: 1,wherein the dispersin variant has hexosaminidase activity preferablybeta-1,6 N-acetylglucosaminidase activity, and wherein the variant hasincreased stability, preferably thermo stability measured as half-lifeimprovement factor, HIF, compared to the dispersin with SEQ ID NO: 1

One aspect of the invention relates to a dispersin variant, comprisingan alteration, preferably substitution at one or more positionscorresponding to positions 2, 3, 12, 15, 17, 18, 19, 22, 23, 24, 25, 26,30, 32, 34, 43, 44, 45, 49, 52, 54, 56, 57, 58, 59, 60, 62, 63, 67, 68,71, 72, 74, 77, 79, 80, 81, 82, 90, 99, 100, 103, 104, 105, 106, 107,110, 111, 113, 114, 116, 117, 118, 119, 120, 122, 123, 124, 125, 126,127, 128, 131, 135, 138, 139, 140, 142, 145, 147, 148, 149, 150, 151,152, 163, 164, 167, 168, 170, 171, 173, 174, 175, 177, 178, 179, 181,185, 186, 187, 188, 189, 199, 200, 203, 204, 205, 207, 208, 210, 215,217, 218, 221, 222, 224, 225, 227, 230, 232, 233, 234, 235, 237, 244,249, 251, 252, 253, 254, 256, 260, 261, 262, 263, 264, 265, 267, 268,270, 271, 272, 273, 274, 276, 278, 279, 280, 281, 282, 283, 284, 287,288, 290, 291, 296, 300, 301, 303, 304, 305, 306, 308, 309, 312, 314,315, 319, 321 and 323 of the polypeptide of SEQ ID NO: 1, wherein thedispersin variant has beta-1,6 N-acetylglucosaminidase activity, whereinthe dispersin variant has at least 80%, at least 85%, at least 90%, atleast 95%, at least 96%, at least 97%, at least 98%, or at least 99%,but less than 100% sequence identity to the polypeptide shown in SEQ IDNO: 1, and wherein the variant has increased stability, preferablythermo stability measured as half-life improvement factor, HIF, comparedto the dispersin of SEQ ID NO: 1.

One aspect of the invention relates to a dispersin variant, comprisingat least one alteration selected from the group consisting of: D2A, D2L,D2N, D2R, D2V, D2W, Q3F, Q3I, Q3L, Q3M, Q3P, Q3V, Q3Y, Q3T, S12A, H15F,H15Y, T17W, T17C, T17E, T17F, T17M, T17R, T17V, V18L, E19D, E19N, E19P,K22A, K22M, K22V, S23C, S23E, S23I, S23L, S23R, S23T, S23V, V25R, D26M,Y30*, Y30D, Y30L, Y30M, Y30N, Y30R, Y30T, Y30V, G32L, G32M, G32R, N43*,N43H, N43L, E44*, N45D, N45L, N45V, A49W, A49Y, Y52*, Y52M, G54L, G54M,G54N, S56T, S56W, S57W, E58N, N59A, N59C, N59D, N59E, N59F, N59M, N59R,N59V, N59W, T60V, N62C, N62D, N62H, N62Q, N62W, T63C, T63D, T63L, T63N,T63R, T63V, K67A, K67L, N68L, N68Q, L71H, L71N, L71R, L71V, L71W, S72*,S72C, S72D, S72E, S72F, S72G, S72I, S72M, S72N, S72R, S72T, S72Y, I74L,S77A, D79V, K80*, K80E, K80H, K80L, K80N, K80Q, K80V, K80W, D81A, D81G,D81L, D81R, D81S, D81T, D81V, D81W, I82V, L90F, E99Q, E99R, L100S,K103A, K103R, K104N, K104W, D105N, V106A, V106D, V106E, V106H, V106K,V106L, V106M, V106N, V106Q, V106W, V106Y, K107A, K107C, K107L, K107M,K107T, K107V, K107W, N110M, N110R, N110V, D111A, D111E, D111M, D111N,D111Q, D111R, D111V, D111W, V113T, T114C, T114S, Y116D, Y116N, Y116R,S117*, S117D, S117H, S117N, S117P, E118*, E118A, E118D, E118G, E118L,E119G, E119W, T120I, T120L, T120M, T120V, T120W, D122*, D122H, D122R,Y123W, Y124C, Y124I, Y124K, Y124L, Y124M, Y124Q, Y124R, Y124T, Y124V,Y124W, D125C, D125G, D125K, D125Q, D125R, N126V, R127D, R127H, R127K,R127L, R127M, R127Q, R127W, V128C, V128L, V128T, D131V, Q135*, Q135A,Q135D, Q135E, Q135K, Q135M, Q135Y, D138K, D138L, D138M, D138Q, D138R,D138S, D138V, D138W, E139W, D142R, D142W, Y145*, Y145H, Y145L, Y145N,Y145V, P147A, P147C, P147D, P147F, P147G, P147L, P147M, P147R, P147S,P147T, P147V, K148A, K148D, K148L, K148V, F149L, F149M, F149N, E150D,E150H, E150K, E150L, E150M, E150N, E150R, E150V, E150W, E150Y, G151A,G151C, G151D, G151L, G151N, G151P, G151S, G151W, K152D, K152L, K152R,G164D, G164E, G164H, G164S, G164V, V167D, V167E, V167L, V167P, V167Q,V167R, V167W, H168N, L170A, L170D, L170E, L170F, L170H, L170K, L170M,L170N, L170P, L170Q, L170R, L170S, L170V, L170W, L170Y, D171A, D171C,D171E, D171K, D171L, D171M, D171Q, D171R, D171V, D171W, D171Y, I173C,D174H, D174M, D174N, D174V, D174W, F175Y, N177M, Q178*, Q178A, Q178K,Q178R, Q178W, I179T, S181C, S181D, S181F, S181G, S181N, S181P, S181Q,S181T, S181V, S181W, E185M, E185R, E185V, E185W, S186D, S186E, S186H,S186I, S186K, S186L, S186M, S186N, S186Q, S186R, S186V, S186W, K187C,K187D, K187G, K187R, K187S, K187V, K187W, Y188P, E189L, E189V, E189W,S199C, S199L, S199M, S199Y, E200D, E200F, E200K, E200L, E200M, E200N,E200R, E200W, A203C, A203D, A203E, A203G, A203L, A203M, A203P, A203R,A203S, A203T, A203V, A203W, N204L, N204M, N204V, N204W, N204Y, L205I,D207A, D207C, D207E, D207G, D207N, D207Q, D207S, D207V, D207W, S208A,S208C, S208D, S208G, S208L, S208Q, S208T, S208V, S208W, S210T, Q215R,Q215M, Q215L, Q215*, S217V, T218A, T218L, T218Q, T218R, T218V, G222D,E224A, E224P, N227A, N227Q, N227R, N227S, N227T, N227K, D230*, D230R,D230T, D230W, E232D, E232V, N233H, N233Q, N233R, N233W, W234R, G235W,G235A, G235E, G235F, G235H, G235I, G235L, G235M, G235N, G235P, G235S,G235V, S237C, S237G, S237M, S237N, S237W, S237Y, Y244C, Y244E, Y244M,L249H, L249K, L249Q, L249R, L249W, L249Y, S251L, S251N, S251R, S251W,N252P, N252C, G253D, G253W, F254I, F254L, F254M, F254Y, Q256E, Q256R,N260*, N260A, N260C, N260E, N260I, N260K, N260L, N260M, N260Q, N260R,N260V, N260W, N260Y, E261*, E261A, E261D, E261R, E261W, Q262*, Q262F,Q262H, Q262W, Q262Y, M263K, M263L, M263Q, D264*, D264C, D264E, Y265F,N267S, N267T, W268C, W268E, W268M, W268R, Y270F, A271D, A271G, H272D,H272I, H272M, H272P, H272V, H272W, N273W, K274R, K274A, K274H, F276A,F276C, F276K, F276N, F276G, F276L, F276M, F276P, F276S, F276V, F276W,I278A, I278K, I278N, I278Q, I278V, S279C, S279D, S279E, S279G, S279N,D280C, D280E, Y281*, Y281A, Y281C, Y281H, Y281K, Y281N, Y281P, Y282E,Y282N, Y281R, H283I, A284I, A284L, A284N, A284P, A284T, A284V, T287N,S288D, S288K, S288N, V290I, K291L, K291R, K291V, T296C, E300A, E300D,H301C, H301N, H301R, T303A, T303C, T303G, T303K, T303Q, T303R, T303W,D304C, D304M, L305M, L305N, S306C, K308A, K308D, K308G, K308I, K308L,K308Q, K308S, K308T, K308V, K308Y, K309A, K309C, K309D, K309H, K309L,K309M, K309N, K309Q, K309S, K309T, K309I, K312A, K312L, K312M, K312N,K312Q, K312S, K312W, E314I, E314L, E314V, L315I, L315V, R319A, andN323R, wherein the positions correspond to the positions of thepolypeptide shown in SEQ ID NO: 1, wherein the dispersin variant hashexosaminidase activity preferably beta-1,6 N-acetylglucosaminidaseactivity, preferably wherein the alteration at the one or moreposition(s) produces a dispersin variant having increased stability,preferably thermo-stability measured as improvement factor, HIF, of atleast 1.1 compared to the polypeptide shown in SEQ ID NO: 1 andpreferably wherein dispersin variant has at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, at least 96%, at least 97%, at least 98%, or at least 99%,but less than 100% sequence identity to the polypeptide shown in SEQ IDNO: 1.

One aspect of the invention relates to a method for obtaining adispersin variant, comprising;

-   -   a) introducing into a dispersin parent having hexosaminidase,        preferably beta-1,6-N-acetyl glycosaminidase activity, an        alteration, preferably a substitution at one or more positions        corresponding to positions 2, 3, 12, 15, 17, 18, 19, 22, 23, 24,        25, 26, 30, 32, 34, 43, 44, 45, 49, 52, 54, 56, 57, 58, 59, 60,        62, 63, 67, 68, 71, 72, 74, 77, 79, 80, 81, 82, 90, 99, 100,        103, 104, 105, 106, 107, 110, 111, 113, 114, 116, 117, 118, 119,        120, 122, 123, 124, 125, 126, 127, 128, 131, 135, 138, 139, 140,        142, 145, 147, 148, 149, 150, 151, 152, 163, 164, 167, 168, 170,        171, 173, 174, 175, 177, 178, 179, 181, 185, 186, 187, 188, 189,        199, 200, 203, 204, 205, 207, 208, 210, 215, 217, 218, 221, 222,        224, 225, 227, 230, 232, 233, 234, 235, 237, 244, 249, 251, 252,        253, 254, 256, 260, 261, 262, 263, 264, 265, 267, 268, 270, 271,        272, 273, 274, 276, 278, 279, 280, 281, 282, 283, 284, 287, 288,        290, 291, 296, 300, 301, 303, 304, 305, 306, 308, 309, 312, 314,        315, 319, 321 or 323 of the polypeptide of SEQ ID NO: 1,    -   b) and recovering the variant.

One aspect of the invention relates to a method for obtaining adispersin variant, comprising;

-   -   a) introducing into a dispersin parent having 1,6-N-acetyl        glycosaminidase activity, an alteration, preferably a        substitution at one or more positions corresponding to positions        2, 3, 12, 15, 17, 18, 19, 22, 23, 24, 25, 26, 30, 32, 34, 43,        44, 45, 49, 52, 54, 56, 57, 58, 59, 60, 62, 63, 67, 68, 71, 72,        74, 77, 79, 80, 81, 82, 90, 99, 100, 103, 104, 105, 106, 107,        110, 111, 113, 114, 116, 117, 118, 119, 120, 122, 123, 124, 125,        126, 127, 128, 131, 135, 138, 139, 140, 142, 145, 147, 148, 149,        150, 151, 152, 163, 164, 167, 168, 170, 171, 173, 174, 175, 177,        178, 179, 181, 185, 186, 187, 188, 189, 199, 200, 203, 204, 205,        207, 208, 210, 215, 217, 218, 221, 222, 224, 225, 227, 230, 232,        233, 234, 235, 237, 244, 249, 251, 252, 253, 254, 256, 260, 261,        262, 263, 264, 265, 267, 268, 270, 271, 272, 273, 274, 276, 278,        279, 280, 281, 282, 283, 284, 287, 288, 290, 291, 296, 300, 301,        303, 304, 305, 306, 308, 309, 312, 314, 315, 319, 321 and 323 of        the polypeptide of SEQ ID NO: 1,    -   b) and recovering the variant.

One aspect of the invention relates to a composition, preferably acleaning composition comprising a variant according to the invention.

One aspect of the invention relates to a method of cleaning an itemcomprising a) adding a dispersin variant according to the invention to acleaning composition and b) washing item with the composition, whereinthe item is a textile or a hard surface.

One aspect of the invention relates to a method for finding improvedpolypeptide variants, comprising the steps of:

-   -   a) generate diversity in selected, preferably all, positions of        a polypeptide, thereby preparing libraries of polypeptide        variants (V_(L)), and    -   b) select subset library (V_(LSub)) with improved variants.

The method above preferably comprises subjecting some subset of variantsto one or more stress condition and then leave a reference dispersinvariants unstressed. In that way the subset can be compared, andimproved variants selected.

One aspect of the invention relates to a method for finding improvedpolypeptide variants, comprising the steps of:

-   -   a) generate diversity in selected, preferably all, positions of        a polypeptide, thereby preparing a library of polypeptide        variants (V_(L)),    -   b) expose part of the library of polypeptide variants (V_(L)) to        one or more stress conditions suitable for measuring a property        of interest, thereby generating a stressed V_(LS+n) and an        unstressed V_(LUS) library;    -   c) compare the stressed V_(LS) and the unstressed V_(LUS)        library; and    -   d) select subset library (V_(LSub)) with improved variants.

DEFINITIONS

The term “adjunct materials” means any liquid, solid or gaseous materialselected for the particular type of detergent composition desired andthe form of the product (e.g., liquid, granule, powder, bar, paste,spray, tablet, gel, or foam composition), which materials are alsopreferably compatible with the dispersin variant enzyme used in thecomposition. In some aspects, granular compositions are in “compact”form, while in other aspects, the liquid compositions are in a“concentrated” form.

The term “allelic variant” means any of two or more alternative forms ofa gene occupying the same chromosomal locus. Allelic variation arisesnaturally through mutation and may result in polymorphism withinpopulations. Gene mutations can be silent (no change in the encodedpolypeptide) or may encode polypeptides having altered amino acidsequences. An allelic variant of a polypeptide is a polypeptide encodedby an allelic variant of a gene.

The term “biofilm” means any group of microorganisms in which cellsstick to each other on a surface, such as a textile, dishware or hardsurface. These adherent cells are frequently embedded within aself-produced matrix of extracellular polymeric substance (EPS). BiofilmEPS is a polymeric conglomeration generally composed of extracellularDNA, proteins, and polysaccharides. Biofilms may form on living ornon-living surfaces. The microbial cells growing in a biofilm arephysiologically distinct from planktonic cells of the same organism,which, by contrast, are single-cells that may float or swim in a liquidmedium. Bacteria living in a biofilm usually have significantlydifferent properties from free-floating bacteria of the same species, asthe dense and protected environment of the film allows them to cooperateand interact in various ways. One benefit of this environment isincreased resistance to detergents and antibiotics, as the denseextracellular matrix and the outer layer of cells protect the interiorof the community. On laundry biofilm producing bacteria can be foundamong the following species: Acinetobacter sp., Aeromicrobium sp.,Brevundimonas sp., Microbacterium sp., Micrococcus luteus, Pseudomonassp., Staphylococcus epidermidis, and Stenotrophomonas sp.

The term “biosoil” mean a stain composed of several types of organicmaterial such as protein, starch, grease but also e.g. polysaccharides.

The term “cDNA” means a DNA molecule that can be prepared by reversetranscription from a mature, spliced, mRNA molecule obtained from aprokaryotic or eukaryotic cell. A cDNA lacks intron sequences that maybe present in the corresponding genomic DNA. The initial, primary RNAtranscript is a precursor to mRNA that is processed through a series ofsteps, including splicing, before appearing as mature spliced mRNA.

The term “clade” means a group of polypeptides clustered together basedon homologous features traced to a common ancestor. Polypeptide cladescan be visualized as phylogenetic trees and a clade is a group ofpolypeptides that consists of a common ancestor and all its linealdescendants. Polypeptides forming a group e.g. a clade as shown in aphylogenetic tree may often share common properties and are alsofunctionally more closely related than other polypeptides not in theclade.

The term “coding sequence” means a polynucleotide, which directlyspecifies the amino acid sequence of its polypeptide product. Theboundaries of the coding sequence are generally determined by an openreading frame, which usually begins with the ATG start codon oralternative start codons such as GTG and TTG and ends with a stop codonsuch as TAA, TAG, and TGA. The coding sequence may be a DNA, cDNA,synthetic, or recombinant polynucleotide.

The term “control sequences” means all components necessary for theexpression of a polynucleotide encoding a variant of the presentinvention. Each control sequence may be native or foreign to thepolynucleotide encoding the variant or native or foreign to each other.Such control sequences include, but are not limited to, a leader,polyadenylation sequence, propeptide sequence, promoter, signal peptidesequence, and transcription terminator. At a minimum, the controlsequences include a promoter, and transcriptional and translational stopsignals. The control sequences may be provided with linkers forintroducing specific restriction sites facilitating ligation of thecontrol sequences with the coding region of the polynucleotide encodinga variant.

By the term “deep-cleaning” is meant disruption or removal of componentsof organic matter, e.g. biosoil or biofilm and EPS, such aspolysaccharides, PNAG, proteins, DNA, soil or other components presentin the organic matter.

The term “detergent composition”, includes unless otherwise indicated,granular or powder-form all-purpose or heavy-duty washing agents,especially cleaning detergents; liquid, gel or paste-form all-purposewashing agents, especially the so- called heavy-duty liquid (HDL) types;liquid fine-fabric detergents; hand dishwashing agents or light dutydishwashing agents, especially those of the high-foaming type; machinedishwashing agents, including the various tablet, granular, liquid andrinse-aid types for household and institutional use; liquid cleaning anddisinfecting agents, including antibacterial hand-wash types, cleaningbars, soap bars, mouthwashes, denture cleaners, car or carpet shampoos,bathroom cleaners; hair shampoos and hair-rinses; shower gels, foambaths; metal cleaners; as well as cleaning auxiliaries such as bleachadditives and “stain-stick” or pre-treat types. The terms “detergentcomposition” and “detergent formulation” are used in reference tomixtures which are intended for use in a wash medium for the cleaning ofsoiled objects. In some aspects, the term is used in reference tolaundering fabrics and/or garments (e.g., “laundry detergents”). Inalternative aspects, the term refers to other detergents, such as thoseused to clean dishes, cutlery, etc. (e.g., “dishwashing detergents”). Itis not intended that the present invention be limited to any particulardetergent formulation or composition. The term “detergent composition”is not intended to be limited to compositions that contain surfactants.It is intended that in addition to the variants according to theinvention, the term encompasses detergents that may contain, e.g.,surfactants, builders, chelators or chelating agents, bleach system orbleach components, polymers, fabric conditioners, foam boosters, sudssuppressors, dyes, perfume, tannish inhibitors, optical brighteners,bactericides, fungicides, soil suspending agents, anti-corrosion agents,enzyme inhibitors or stabilizers, enzyme activators, transferase(s),hydrolytic enzymes, oxido reductases, bluing agents and fluorescentdyes, antioxidants, and solubilizers.

The term “dispersin” and the abbreviations “Dsp” or “Disp” arepolypeptides having hexosaminidase activity, EC 3.2.1.—that catalyzesthe hydrolysis of β-1,6-glycosidic linkages of N-acetyl-glucosaminepolymers (poly-N-acetylglucosamine, PNAG) found e.g. in biofilm, EPS,cell debris and other biosoils. Thus, dispersins are enzymes havingbeta-1,6 N-acetylglucosaminidase activity orpoly-beta-1,6-N-actylglucosamin (PNAG) activity. For purposes of thepresent invention, dispersin activity i.e. beta-1,6N-acetylglucosaminidase activity is determined according to theprocedure described in the activity assay of Example 3. In some aspects,the dispersin variants of the present invention have improved dispersinactivity compared to the parent dispersin. In some aspects, thedispersin variants of the present invention have at least 100%, e.g., atleast 110%, at least 120%, at least 130%, at least 140%, at least 150%,at least 160%, at least 170%, at least 180%, at least 190%, or at least200% dispersin activity compared to the polypeptide shown in SEQ ID NO:1 or such as from 100% to 200% dispersin activity compared to thepolypeptide shown in SEQ ID NO: 1.

The term “effective amount of enzyme” refers to the quantity of enzymenecessary to achieve the enzymatic activity required in the specificapplication, e.g., in a defined detergent composition. Such effectiveamounts are readily ascertained by one of ordinary skill in the art andare based on many factors, such as the particular enzyme used, thecleaning application, the specific composition of the detergentcomposition, and whether a liquid or dry (e.g., granular, bar)composition is required, and the like. The term “effective amount” of adispersin variant refers to the quantity of dispersin variant describedhereinbefore that achieves a desired level of enzymatic activity, e.g.,in a defined detergent composition.

The term “expression” includes any step involved in the production ofthe variant including, but not limited to, transcription,post-transcriptional modification, translation, post-translationalmodification, and secretion.

The term “expression vector” means a linear or circular DNA moleculethat comprises a polynucleotide encoding a variant and is operablylinked to additional nucleotides that provide for its expression.

The term “fabric” encompasses any textile material. Thus, it is intendedthat the term encompass garments, as well as fabrics, yarns, fibers,non-woven materials, natural materials, synthetic materials, and anyother textile material.

The term “high detergent concentration” system includes detergentswherein greater than about 2000 ppm of detergent components is presentin the wash water. European detergents are generally considered to behigh detergent concentration systems.

The term “host cell” means any cell type that is susceptible totransformation, transfection, transduction, and the like with a nucleicacid construct or expression vector comprising a polynucleotide of thepresent invention. The term “host cell” encompasses any progeny of aparent cell that is not identical to the parent cell due to mutationsthat occur during replication.

The term “improved property” means a characteristic associated with avariant that is improved compared to the parent and/or compared to adispersin with SEQ ID NO: 1 or compared to a dispersin having theidentical amino acid sequence of said variant but not having thealterations at two or more of said specified positions. Such improvedproperties include, but are not limited to, stability, such as detergentstability, wash performance e.g. deep cleaning effect and thedeep-cleaning effect may include but is not limited to de-gluing effect(some organic materials such as EPS is sticky i.e. glue-like and coursesdirt to stick to the material), improved whiteness, reduction of malodorand re-deposition. Improved property also includes improved stability inpresence of certain enzymes e.g. proteases (protease stability) orsubstances e.g. stability in presence of sulfite e.g. sulfite stability.

The term “improved dispersin activity” is defined herein as an altereddispersin activity e.g. by increased catalyse of β-1,6-glycosidiclinkages of N-acetyl-glucosamine polymers (poly-N-acetylglucosamine,PNAG) i.e. the dispersin variant displaying an alteration of theactivity relative (or compared) to the activity of the parent dispersin,such as compared to a dispersin with SEQ ID NO: 1 or compared to adispersin having the identical amino acid sequence of said variant butnot having the alterations at one or more of said specified positions.

The term “low detergent concentration” system includes detergents whereless than about 800 ppm of detergent components is present in the washwater. Asian, e.g., Japanese detergents are typically considered lowdetergent concentration systems.

The term “improved wash performance” includes but is not limited to theterm “deep cleaning effect”. Improved performance e.g. deep cleaningperformance of a dispersin variant according to the invention ismeasured compared to the dispersin parent e.g. the dispersin shown inSEQ ID NO: 1 or compared to a dispersin having the identical amino acidsequence of said variant but not having the alterations at one or moreof said specified positions. The improved performance e.g. deep cleaningperformance may be expressed as a Remission value of the stainedswatches. After washing and rinsing the swatches are spread out flat andallowed to air dry at room temperature overnight. All washed swatchesare evaluated the day after the wash. Light reflectance evaluations ofthe swatches are done using a Macbeth Color Eye 7000 reflectancespectrophotometer with very small aperture. The measurements are madewithout UV in the incident light and remission at 460 nm is extracted.Positive responses indicate that soil is removed.

The term “laundering” relates to both household laundering andindustrial laundering and means the process of treating textiles with asolution containing a cleaning or detergent composition of the presentinvention. The laundering process can for example be carried out usinge.g. a household or an industrial washing machine or can be carried outby hand.

The term “mature polypeptide” means a polypeptide in its final formfollowing translation and any post-translational modifications, such asN-terminal processing, C-terminal truncation, glycosylation,phosphorylation, etc. In some aspects, the mature polypeptide is aminoacids 1 to 324 of SEQ ID NO: 1. The N-terminals of the maturepolypeptide used according to the present invention may beexperimentally confirmed based on EDMAN N-terminal sequencing data andIntact MS data. It is known in the art that a host cell may produce amixture of two of more different mature polypeptides (i.e., with adifferent C-terminal and/or N-terminal amino acid) expressed by the samepolynucleotide.

The term “medium detergent concentration” system includes detergentswherein between about 800 ppm and about 2000 ppm of detergent componentsis present in the wash water. North American detergents are generallyconsidered to be medium detergent concentration systems.

The term “nucleic acid construct” means a nucleic acid molecule, eithersingle- or double-stranded, which is isolated from a naturally occurringgene or is modified to contain segments of nucleic acids in a mannerthat would not otherwise exist in nature or which is synthetic. The termnucleic acid construct is synonymous with the term “expression cassette”when the nucleic acid construct contains the control sequences requiredfor expression of a coding sequence of the present invention.

The term “non-fabric detergent compositions” include non-textile surfacedetergent compositions, including but not limited to compositions forhard surface cleaning, such as dishwashing detergent compositionsincluding manual dish wash compositions, oral compositions, denturecompositions, and personal cleansing compositions.

The term “operably linked” means a configuration in which a controlsequence is placed at an appropriate position relative to the codingsequence of a polynucleotide such that the control sequence directs theexpression of the coding sequence.

The term “parent” dispersin, dispersin parent or precursor dispersin maybe used interchangeably. In context of the present invention “parentdispersin” is to be understood as a dispersin into which at least onealteration is made in the amino acid sequence to produce a dispersinvariant having an amino acid sequence which is less than 100% identicalto the dispersin sequence into which the alteration was made i.e. theparent dispersin. Thus, the parent is a dispersin having identical aminoacid sequence compared to the variant but not having the alterations atone or more of the specified positions. It will be understood, that inthe present context the expression “having identical amino acidsequence” relates to 100% sequence identity. In a particular aspect thedispersin parent is a dispersin having at least 60%, at least 61%, atleast 62%, at least 63%, at least 64%, at least 65%, at least 66%, atleast 67%, at least 68%, at least 69%, at least 70%, at least 72%, atleast 73%, at least 74%, at least 75%, at least 80%, at least 81%, atleast 82%, at least 83%, at least 84%, at least 85%, at least 90%, atleast 91%, at least 92%, at least 93%, at least 94%, at least 95%, atleast 96%, at least 97%, at least 98%, at least 99%, e. g. at least99.1%, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%,at least 99.6 or 100% identity to a polypeptide shown in SEQ ID NO: 1.

The term “relevant washing conditions” is used herein to indicate theconditions, particularly washing temperature, time, washing mechanics,detergent concentration, type of detergent and water hardness, actuallyused in households in a detergent market segment.

The term “stability” includes storage stability and stability duringuse, e.g. during a wash process and reflects the stability of thedispersin variant according to the invention as a function of time e.g.how much activity is retained when the dispersin variant is kept insolution in particular in a detergent solution. The stability isinfluenced by many factors e.g. pH, temperature, detergent compositione.g. amounts of builder, surfactants etc. The dispersin stability may bemeasured as improved RA (residual activity), T_(m) (melting temperature)or T_(1/2) (half-life) as described in Examples 2a, 2b, 3, 4, 5 and 6.The term “improved stability” or “increased stability” is defined hereinas a variant dispersin displaying an increased stability in solutions,relative to the stability of the parent dispersin and/or relative to SEQID NO: 1. “Improved stability” and “increased stability” includesdetergent stability. The term “detergent stability” or “improveddetergent stability is defined as improved stability in a detergent(e.g. cleaning composition) compared to the parent dispersin. Thedispersin variant improvement may be measured by RAR (residual activityratio), HIF (Half-life improvement factor) o T_(m), described in Example2a, 2b, 3, 4, 5 and 6.

The term “textile” refers to woven fabrics, as well as staple fibers andfilaments suitable for conversion to or use as yarns, woven, knit, andnon-woven fabrics. The term encompasses yarns made from natural, as wellas synthetic (e.g., manufactured) fibers. The term, “textile materials”is a general term for fibers, yarn intermediates, yarn, fabrics, andproducts made from fabrics (e.g., garments and other articles).

The term “transcription terminator” is used for a section of the geneticsequence that marks the end of gene or operon on genomic DNA fortranscription.

The term “variant” means a polypeptide which comprises an alteration atone or more (e.g., several) positions compared to the parent orreference polypeptide. The alteration may be a substitution, insertionor deletion. A substitution means replacement of the amino acidoccupying a position with a different amino acid, a deletion meansremoval of an amino acid occupying a position and an insertion meansadding amino acids e.g. 1 to 10 amino acids, preferably 1-3 amino acidsadjacent to an amino acid occupying a position. The term “dispersinvariant” means a polypeptide having hexosaminidase, preferably beta-1,6N-acetylglucosaminidase activity or is activetopoly-beta-1,6-N-actylglucosamin (PNAG) and which comprise analteration, i.e., a substitution, insertion, and/or deletion at one ormore (or one or several) positions compared to the parent dispersin e.g.compared to SEQ ID NO: 1. The term “dispersin activity” means apolypeptide having hexosaminidase, preferably beta-1,6N-acetylglucosaminidase activity or is activetopoly-beta-1,6-N-actylglucosamin (PNAG). The variants e.g. dispersinvariants of the present invention preferably have at least 20%, e.g., atleast 40%, at least 50%, at least 60%, at least 70%, at least 80%, atleast 90%, at least 95%, or at least 100% of the dispersin activity ofthe polypeptide shown in SEQ ID NO: 1.

The term “water hardness” or “degree of hardness” or “dH” or “° dH” asused herein refers to German degrees of hardness. One degree is definedas 10 milligrams of calcium oxide per liter of water.

The term “wild-type dispersin” means a dispersin expressed by anaturally occurring organism, such as a fungal, bacterium, archaea,yeast, plant or animal found in nature.

Conventions for Designation of Variants

For purposes of the present invention, the polypeptide disclosed in SEQID NO: 1 is used to determine the corresponding amino acid residue inanother dispersin. The amino acid sequence of another dispersin isaligned with the polypeptide disclosed in SEQ ID NO: 1, and based on thealignment, the amino acid position number corresponding to any aminoacid residue in the polypeptide disclosed in SEQ ID NO: 1 is determinedusing e.g. the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970,J. Mol. Biol. 48: 443-453) as implemented in the Needle program of theEMBOSS package (EMBOSS: The European Molecular Biology Open SoftwareSuite, Rice et al., 2000, Trends Genet. 16: 276-277), preferably version5.0.0 or later. The parameters used are gap open penalty of 10, gapextension penalty of 0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM62)substitution matrix.

Identification of the corresponding amino acid residue in anotherdispersin can be determined by an alignment of multiple polypeptidesequences using several computer programs including, but not limited to,MUSCLE (multiple sequence comparison by log-expectation; version 3.5 orlater; Edgar, 2004, Nucleic Acids Research 32: 1792-1797), MAFFT(version 6.857 or later; Katoh and Kuma, 2002, Nucleic Acids Research30: 3059-3066; Katoh et al., 2005, Nucleic Acids Research 33: 511-518;Katoh and Toh, 2007, Bioinformatics 23: 372-374; Katoh et al., 2009,Methods in Molecular Biology 537:_39-64; Katoh and Toh, 2010,Bioinformatics 26:_1899-1900), and EMBOSS EMMA employing ClustalW (1.83or later; Thompson et al., 1994, Nucleic Acids Research 22: 4673-4680),using their respective default parameters.

When another enzyme has diverged from e.g. the polypeptide of SEQ ID NO:1 such that traditional sequence-based comparison fails to detect theirrelationship (Lindahl and Elofsson, 2000, J. Mol. Biol. 295: 613-615),other pairwise sequence comparison algorithms can be used. Greatersensitivity in sequence-based searching can be attained using searchprograms that utilize probabilistic representations of polypeptidefamilies (profiles) to search databases. For example, the PSI-BLASTprogram generates profiles through an iterative database search processand is capable of detecting remote homologs (Atschul et al., 1997,Nucleic Acids Res. 25: 3389-3402). Even greater sensitivity can beachieved if the family or superfamily for the polypeptide has one ormore representatives in the protein structure databases. Programs suchas GenTHREADER (Jones, 1999, J. Mol. Biol. 287: 797-815; McGuffin andJones, 2003, Bioinformatics 19: 874-881) utilize information from avariety of sources (PSI-BLAST, secondary structure prediction,structural alignment profiles, and solvation potentials) as input to aneural network that predicts the structural fold for a query sequence.Similarly, the method of Gough et al., 2000, J. Mol. Biol. 313: 903-919,can be used to align a sequence of unknown structure with thesuperfamily models present in the SCOP database. These alignments can inturn be used to generate homology models for the polypeptide, and suchmodels can be assessed for accuracy using a variety of tools developedfor that purpose.

For proteins of known structure, several tools and resources areavailable for retrieving and generating structural alignments. Forexample, the SCOP super families of proteins have been structurallyaligned, and those alignments are accessible and downloadable. Two ormore protein structures can be aligned using a variety of algorithmssuch as the distance alignment matrix (Holm and Sander, 1998, Proteins33: 88-96) or combinatorial extension (Shindyalov and Bourne, 1998,Protein Engineering 11: 739-747), and implementation of these algorithmscan additionally be utilized to query structure databases with astructure of interest in order to discover possible structural homologs(e.g., Holm and Park, 2000, Bioinformatics 16: 566-567).

As different amino acids may be present at a given position depending onthe selected parent for the variants the amino acid positions areindicated with #₁, #₂, etc. in the definitions below. In describing thevariants of the present invention, the nomenclature described below isadapted for ease of reference. The accepted IUPAC single letter or threeletters amino acid abbreviations are employed.

Substitutions

For an amino acid substitution, the following nomenclature is used:Original amino acid, position, substituted amino acid. Accordingly, thesubstitution of valine at position #₁ with alanine is designated as “Val#₁Ala” or “V #₁A”. Multiple mutations may be separated by addition marks(“+”) or by commas (,), e.g., “Val #₁Ala+“Pro #₂Gly” or V #₁A, P #₂G,representing substitutions at positions #₁ and #₂ of valine (V) andproline (P) with alanine (A) and glycine (G), respectively. If more thanone amino acid may be substituted in a given position these may belisted in brackets, such as [X] or {X}. Thus, if both Trp and Lys may besubstituted instead of the amino acid occupying at position #₁ this maybe indicated as X #₁ {W, K}, X #₁ [W, K] or X #₁ [W/K], where the Xindicate the amino acid residue present at the position of the parentdispersin e.g. such as a dispersin shown in SEQ ID NO: 1 or a dispersinhaving at least 60% identity hereto. In some cases, the variants may berepresented as #₁ {W, K} or X #₂P emphasizing that the amino acids to besubstituted vary depending on the parent. For convenience, as SEQ ID NO:1 is used for numbering, the amino acid in the corresponding position inSEQ ID NO: 1 is indicated, e.g. D2V. However, it will be clear to theskilled artisan that a dispersin variant comprising D2V is not limitedto parent dispersins having a “D” (aspartic acid) at a positioncorresponding to position 2 of SEQ ID NO: 1. In a parent dispersinhaving e.g. asparagine in position 2, the skilled person would translatethe mutation specified as D2V to N2V. In the event the parent dispersinhas valine in position 1, the skilled person would recognize that theparent dispersin is not changed at this position. The same applies fordeletions and insertions described below.

Deletions

For an amino acid deletion, the following nomenclature is used: Originalamino acid, position, *. Accordingly, the deletion of valine at position#₁ is designated as “Val #₁*” or “V #₁*”. Multiple deletions may beseparated by addition marks (“+”) or commas, e.g., “Val #₁*+Pro #₂*” or“V #₁*, P #₂*”.

Insertions

The insertion of an additional amino acid residue such as e.g. a lysineafter Val #₁ may be indicated by: Val #₁ValLys or V #₁VK. Alternatively,insertion of an additional amino acid residue such as lysine after V #₁may be indicated by: *#₁aK. When more than one amino acid residue isinserted, such as e.g. a Lys, and Gly after #₁ this may be indicated as:Val #₁ValLysGly or V #₁VKG. In such cases, the inserted amino acidresidue(s) may also be numbered by the addition of lower case letters tothe position number of the amino acid residue preceding the insertedamino acid residue(s), in this example: *#₁aK *#₁bG.

Multiple Alterations

Variants comprising multiple alterations may be separated by additionmarks (“+”) or by commas (,), e.g., “Val #₁Trp+Pro #₂Gly” or “V #₁W, P#₂G” representing a substitution of valine and proline at positions #₁and #₂ with tryptophan and glycine, respectively as described above.

Different Alterations

Where different alterations can be introduced at a position, thedifferent alterations may be separated by a comma, e.g., “Val #₁Trp,Lys” or V #₁W, K representing a substitution of valine at position #₁with tryptophan or lysine. Thus, “Val #₁Trp, Lys+Pro #₂Asp” designatesthe following variants: “Val #₁Trp+Pro #₂Asp”, “Val #₁Lys+Pro #₂Asp” orV #₁W, K+P #₂D. The term “substitution set” is in the context of thepresent invention a variant comprising more than one mutation comparedto the parent or reference enzyme e.g. the substitution set D2V+Q3L(compared to SEQ ID NO: 1) is a variant of SEQ ID NO: 1 comprising thetwo mutations D2V+Q3L compared to SEQ ID NO: 1.

When mutations in alteration e.g. substitution sets are separated bycomas or plusses “+” this means that all the alterations in the set arepresent and the selection is between the lists of alterations(alteration sets). E.g. “comprises one or more of the followingsubstitution sets: Q3I+A49W, Q3I+N59E, Q3I+S163P, Q3I+S186R, Q3I+Q215K,Q3I+S225G, Q3I+N227T, Q3I+N252P, Q3I+N267T, Q3I+F276A, Q3I+Y281P,Q3I+K308Q, Q3I+K308E, Q3I+K309E, Q3I+K312Q, Q3I+K312E. . . .

means the selection is made between each of the substitution setsseparated by commas.

The terms parent enzyme includes terms such as reference enzyme, backbone or starting enzyme and is used to denote the enzyme into which tomutations e.g. substitutions are made. The terms may be usedinterchangeably

Specific for Nomenclature of Clades

For purposes of the present invention, the nomenclature [IV] or [I/V]means that the amino acid at this position may be isoleucine (Ile, I) orvaline (Val, V). Likewise, the nomenclature [LVI] and [L/V/I] means thatthe amino acid at this position may be a leucine (Leu, L), valine (Val,V) or isoleucine (Ile, I), and so forth for other combinations asdescribed herein. Unless otherwise limited further, the amino acid X isdefined such that it may be any of the 20 naturally-occurring aminoacids.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides novel dispersins preferably obtained fromTerribacillus, in particular, Terribacillus saccharophilus. Thedispersins of the invention preferably comprise at least 60% sequenceidentity to a polypeptide with SEQ ID NO: 1 and comprise an alterationof at least one amino acid positions compared to the dispersin with SEQID NO: 1. In some aspects, a dispersin variant of the invention comprisean amino acid sequence comprising at least two substitutions of an aminoacid made at positions equivalent to the positions in SEQ ID NO: 1. Thepresent invention also relates to methods for producing dispersinvariants. The dispersin variants of the present invention preferablyhave at least one improved property compared to the parent dispersin orcompared to SEQ ID NO: 1. Properties include but are not limited to:stability such as; stability in detergents, storage stability, in washstability and thermo-stability, wash performance in particulardeep-cleaning performance, increased expression level and malodorreduction. Preferably the improved property is improved stability e.g.stability in detergent (e.g. cleaning compositions). When household caresurfaces such as textiles are soiled the stain is often a complexmixture of various organic matters such as skin debris e.g. dead cells,sweat and sebum, pollution from outside and from e.g. the body. Suchbiosoil composes different organic stains, such as protein, starch,grease but also e.g. polysaccharides. Complex stains, which are oftenstubborn and difficult to remove. The use of dispersins in laundryprocesses and in detergents has been described in WO2017/186943(Novozymes A/S), which disclose the dispersins from Terribacillus suchas dispersins with SEQ ID NO: 1. Dispersins have shown very effective inremoving e.g. PNAG (Poly-N-Acetyl Glucosamine) related stains fromtextiles under laundry conditions. However, when enzymes are formulatedinto cleaning compositions, such as detergents, they may be less activeover time, particularly in liquid detergents. Obviously, its preferablethat enzymes are stable in cleaning and detergent compositions duringstorage (storage stability) and during the wash process (in-washstability). The former may be measured in accelerated stability testswhere the dispersin variants are tested under stressed (e.g. increasedheating) and non-stressed conditions, residual activity (RA) orHalf-life (T_(1/2)) are measured and compared to the values of theparent (or another reference). The residual activity ratio (RAR) orHalf-life improvement factor (HIF) of the reference or parent dispersinmay be assigned the value 1. Thus, dispersin variants having RAR or HIFabove 1 have improved stability under the measured conditions comparedto the parent or reference dispersin. In a preferred embodiment of theinvention the dispersin variants have improved stability compared to theparent dispersin and are therefore advantageous over state-of-the-artdispersins.

Dispersin Variants

Some aspects of the invention relate to dispersin variants of SEQ ID NO:1 or variants of a dispersin having at least 60% identity hereto and tomethods for production of a dispersin variant of SEQ ID NO: 1 or adispersin having at least 60% identity hereto.

In one aspect the present invention relates to a dispersin comprising analteration at one or more position selected from the list consisting ofpositions 2, 3, 12, 15, 17, 18, 19, 22, 23, 24, 25, 26, 30, 32, 34, 43,44, 45, 49, 52, 54, 56, 57, 58, 59, 60, 62, 63, 67, 68, 71, 72, 74, 77,79, 80, 81, 82, 90, 99, 100, 103, 104, 105, 106, 107, 110, 111, 113,114, 116, 117, 118, 119, 120, 122, 123, 124, 125, 126, 127, 128, 131,135, 138, 139, 140, 142, 145, 147, 148, 149, 150, 151, 152, 163, 164,167, 168, 170, 171, 173, 174, 175, 177, 178, 179, 181, 185, 186, 187,188, 189, 199, 200, 203, 204, 205, 207, 208, 210, 215, 217, 218, 221,222, 224, 225, 227, 230, 232, 233, 234, 235, 237, 244, 249, 251, 252,253, 254, 256, 260, 261, 262, 263, 264, 265, 267, 268, 270, 271, 272,273, 274, 276, 278, 279, 280, 281, 282, 283, 284, 287, 288, 290, 291,296, 300, 301, 303, 304, 305, 306, 308, 309, 312, 314, 315, 319, 321 and323, wherein each position corresponds to the position of thepolypeptide of SEQ ID NO: 1, wherein the variant has dispersin activityand preferably wherein the variant has improved stability compared tothe parent e.g. compared to SEQ ID NO: 1.

In one aspect the present invention relates to a dispersin comprising analteration at one or more position selected from the list consisting ofpositions 31, 33, 36, 73, 75, 94, 141, 144, 241, 277, 294, 297, 299,wherein each position corresponds to the position of the polypeptide ofSEQ ID NO: 1, wherein the variant has dispersin activity and preferablywherein the variant has improved stability compared to the parent e.g.compared to SEQ ID NO: 1.

In one preferred aspect the present invention relates to a dispersincomprising an alteration at one or more positions selected from the listconsisting of positions 2, 3, 12, 15, 18, 22, 23, 24, 25, 30, 49, 56,57, 59, 62, 63, 68, 72, 74, 77, 82, 90, 99, 100, 106, 114, 123, 124,125, 135, 138, 163, 167, 170, 171, 173, 174, 175, 178, 179, 181, 185,186, 187, 188, 189, 199, 203, 204, 205, 207, 210, 215, 221, 225, 227,232, 235, 244, 252, 256, 260, 262, 263, 264, 265, 267, 270, 272, 273,274, 276, 278, 279, 280, 281, 282, 283, 284, 288, 290, 291, 296, 303,304, 305, 306, 308, 309, 312, 314, 315, 319, 321, 322 and 323, whereineach position corresponds to the position of the polypeptide of SEQ IDNO: 1, wherein the variant has dispersin activity and preferably whereinthe variant has improved stability compared to the parent e.g. comparedto SEQ ID NO: 1.

In one preferred aspect the present invention relates to a dispersincomprising an alteration at one or more positions selected from the listconsisting of positions 3, 15, 49, 59, 140, 163, 186, 207, 215, 218,225, 227, 232, 235, 237, 252, 260, 267, 272, 276, 279, 281, 288, 308,309 and 312, wherein each position corresponds to the position of thepolypeptide of SEQ ID NO: 1, wherein the variant has dispersin activityand preferably wherein the variant has improved stability compared tothe parent e.g. compared to SEQ ID NO: 1.

In one preferred aspect the present invention relates to a dispersincomprising a substitution at one or more positions selected from thelist consisting of positions 3, 15, 49, 59, 163, 186, 225, 227, 232,235, 252, 260, 272, 279, 281, 308, 309 and 312, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereinthe variant has dispersin activity and preferably wherein the varianthas improved stability compared to the parent e.g. compared to SEQ IDNO: 1.

In one preferred aspect the present invention relates to a dispersincomprising a substitution at two or more positions selected from thelist consisting of positions 3, 15, 49, 59, 163, 186, 225, 227, 232,235, 252, 260, 272, 279, 281, 308, 309 and 312, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereinthe variant has dispersin activity and preferably wherein the varianthas improved stability compared to the parent e.g. compared to SEQ IDNO: 1.

In one preferred aspect the present invention relates to a dispersincomprising a substitution at three or more positions selected from thelist consisting of positions 3, 15, 49, 59, 163, 186, 225, 227, 232,235, 252, 260, 272, 279, 281, 308, 309 and 312, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereinthe variant has dispersin activity and preferably wherein the varianthas improved stability compared to the parent e.g. compared to SEQ IDNO: 1.

In one preferred aspect the present invention relates to a dispersincomprising a substitution at four or more positions selected from thelist consisting of positions 3, 15, 49, 59, 163, 186, 225, 227, 232,235, 252, 260, 272, 279, 281, 308, 309 and 312, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereinthe variant has dispersin activity and preferably wherein the varianthas improved stability compared to the parent e.g. compared to SEQ IDNO: 1.

In one preferred aspect the present invention relates to a dispersincomprising a substitution at five or more positions selected from thelist consisting of positions 3, 15, 49, 59, 163, 186, 225, 227, 232,235, 252, 260, 272, 279, 281, 308, 309 and 312, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereinthe variant has dispersin activity and preferably wherein the varianthas improved stability compared to the parent e.g. compared to SEQ IDNO: 1.

In one preferred aspect the present invention relates to a dispersincomprising a substitution at six or more positions selected from thelist consisting of positions 3, 15, 49, 59, 163, 186, 225, 227, 232,235, 252, 260, 272, 279, 281, 308, 309 and 312, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereinthe variant has dispersin activity and preferably wherein the varianthas improved stability compared to the parent e.g. compared to SEQ IDNO: 1.

In one preferred aspect the present invention relates to a dispersincomprising a substitution at seven or more positions selected from thelist consisting of positions 3, 15, 49, 59, 163, 186, 225, 227, 232,235, 252, 260, 272, 279, 281, 308, 309 and 312, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereinthe variant has dispersin activity and preferably wherein the varianthas improved stability compared to the parent e.g. compared to SEQ IDNO: 1.

In one preferred aspect the present invention relates to a dispersincomprising a substitution at eight or more positions selected from thelist consisting of positions 3, 15, 49, 59, 163, 186, 225, 227, 232,235, 252, 260, 272, 279, 281, 308, 309 and 312, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereinthe variant has dispersin activity and preferably wherein the varianthas improved stability compared to the parent e.g. compared to SEQ IDNO: 1.

In one preferred aspect the present invention relates to a dispersincomprising a substitution at nine or more positions selected from thelist consisting of positions 3, 15, 49, 59, 163, 186, 225, 227, 232,235, 252, 260, 272, 279, 281, 308, 309 and 312, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereinthe variant has dispersin activity and preferably wherein the varianthas improved stability compared to the parent e.g. compared to SEQ IDNO: 1.

In one preferred aspect the present invention relates to a dispersincomprising a substitution at ten or more positions selected from thelist consisting of positions 3, 15, 49, 59, 163, 186, 225, 227, 232,235, 252, 260, 272, 279, 281, 308, 309 and 312, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereinthe variant has dispersin activity and preferably wherein the varianthas improved stability compared to the parent e.g. compared to SEQ IDNO: 1.

In one preferred aspect the present invention relates to a dispersincomprising a substitution at eleven or more positions selected from thelist consisting of positions 3, 15, 49, 59, 163, 186, 225, 227, 232,235, 252, 260, 272, 279, 281, 308, 309 and 312, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereinthe variant has dispersin activity and preferably wherein the varianthas improved stability compared to the parent e.g. compared to SEQ IDNO: 1.

In one preferred aspect the present invention relates to a dispersincomprising a substitution at twelve or more positions selected from thelist consisting of positions 3, 15, 49, 59, 163, 186, 225, 227, 232,235, 252, 260, 272, 279, 281, 308, 309 and 312, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereinthe variant has dispersin activity and preferably wherein the varianthas improved stability compared to the parent e.g. compared to SEQ IDNO: 1.

In one preferred aspect the present invention relates to a dispersincomprising a substitution at thirteen or more positions selected fromthe list consisting of positions 3, 15, 49, 59, 163, 186, 225, 227, 232,235, 252, 260, 272, 279, 281, 308, 309 and 312, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereinthe variant has dispersin activity and preferably wherein the varianthas improved stability compared to the parent e.g. compared to SEQ IDNO: 1.

In one preferred aspect the present invention relates to a dispersincomprising a substitution at fourteen or more positions selected fromthe list consisting of positions 3, 15, 49, 59, 163, 186, 225, 227, 232,235, 252, 260, 272, 279, 281, 308, 309 and 312, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereinthe variant has dispersin activity and preferably wherein the varianthas improved stability compared to the parent e.g. compared to SEQ IDNO: 1.

In one preferred aspect the present invention relates to a dispersincomprising a substitution at fifthteen or more positions selected fromthe list consisting of positions 3, 15, 49, 59, 163, 186, 225, 227, 232,235, 252, 260, 272, 279, 281, 308, 309 and 312, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereinthe variant has dispersin activity and preferably wherein the varianthas improved stability compared to the parent e.g. compared to SEQ IDNO: 1.

In one preferred aspect the present invention relates to a dispersincomprising a substitution at sixteen or more positions selected from thelist consisting of positions 3, 15, 49, 59, 163, 186, 225, 227, 232,235, 252, 260, 272, 279, 281, 308, 309 and 312, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereinthe variant has dispersin activity and preferably wherein the varianthas improved stability compared to the parent e.g. compared to SEQ IDNO: 1.

In one preferred aspect the present invention relates to a dispersincomprising a substitution at seventeen or more positions selected fromthe list consisting of positions 3, 15, 49, 59, 163, 186, 225, 227, 232,235, 252, 260, 272, 279, 281, 308, 309 and 312, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereinthe variant has dispersin activity and preferably wherein the varianthas improved stability compared to the parent e.g. compared to SEQ IDNO: 1.

In one preferred aspect the present invention relates to a dispersincomprising a substitution at all the positions 3, 15, 49, 59, 163, 186,225, 227, 232, 235, 252, 260, 272, 279, 281, 308, 309 and 312, whereineach position corresponds to the position of the polypeptide of SEQ IDNO: 1, wherein the variant has dispersin activity and preferably whereinthe variant has improved stability compared to the parent e.g. comparedto SEQ ID NO: 1.

The term “comprising a substitution” is in the present context meantcomprising a substitution compared to the starting dispersin or theparent. The thus, dispersin have a replacement of the amino acid in e.g.position 3 with another amino acid.

In some aspects the variant has a sequence identity of at least 60%, atleast 65%, at least 70%, at least 75%, at least 80%, at least 85%, atleast 90%, at least 95%, such as at least 96%, at least 97%, at least98%, or at least 99%, but less than 100%, to the amino acid sequence ofthe mature parent dispersin e.g. the dispersin shown in SEQ ID NO: 1.

In some preferred aspects, the dispersin variant comprises an alterationselected from the group consisting of: D2A, D2L, D2N, D2R, D2V, D2W,Q3F, Q3I, Q3L, Q3M, Q3P, Q3V, Q3Y, Q3T, S12A, H15F, H15Y, T17W, T17C,T17E, T17F, T17M, T17R, T17V, V18L, E19D, E19N, E19P, K22A, K22M, K22V,S23C, S23E, S23I, S23L, S23R, S23T, S23V, V25R, D26M, Y30*, Y30D, Y30L,Y30M, Y30N, Y30R, Y30T, Y30V, G32L, G32M, G32R, N43*, N43H, N43L, E44*,N45D, N45L, N45V, A49W, A49Y, Y52*, Y52M, G54L, G54M, G54N, S56T, S56W,S57W, E58N, N59A, N59C, N59D, N59E, N59F, N59M, N59R, N59V, N59W, T60V,N62C, N62D, N62H, N62Q, N62W, T63C, T63D, T63L, T63N, T63R, T63V, K67A,K67L, N68L, N68Q, L71H, L71N, L71R, L71V, L71W, S72*, S72C, S72D, S72E,S72F, S72G, S72I, S72M, S72N, S72R, S72T, S72Y, I74L, S77A, D79V, K80*,K80E, K80H, K80L, K80N, K80Q, K80V, K80W, D81A, D81G, D81L, D81R, D81S,D81T, D81V, D81W, I82V, L90F, E99Q, E99R, L100S, K103A, K103R, K104N,K104W, D105N, V106A, V106D, V106E, V106H, V106K, V106L, V106M, V106N,V106Q, V106W, V106Y, K107A, K107C, K107L, K107M, K107T, K107V, K107W,N110M, N110R, N110V, D111A, D111E, D111M, D111N, D111Q, D111R, D111V,D111W, V113T, T114C, T114S, Y116D, Y116N, Y116R, S117*, S117D, S117H,S117N, S117P, E118*, E118A, E118D, E118G, E118L, E119G, E119W, T120I,T120L, T120M, T120V, T120W, D122*, D122H, D122R, Y123W, Y124C, Y124I,Y124K, Y124L, Y124M, Y124Q, Y124R, Y124T, Y124V, Y124W, D125C, D125G,D125K, D125Q, D125R, N126V, R127D, R127H, R127K, R127L, R127M, R127Q,R127W, V128C, V128L, V128T, D131V, Q135*, Q135A, Q135D, Q135E, Q135K,Q135M, Q135Y, D138K, D138L, D138M, D138Q, D138R, D138S, D138V, D138W,E139W, D142R, D142W, Y145*, Y145H, Y145L, Y145N, Y145V, P147A, P147C,P147D, P147F, P147G, P147L, P147M, P147R, P147S, P147T, P147V, K148A,K148D, K148L, K148V, F149L, F149M, F149N, E150D, E150H, E150K, E150L,E150M, E150N, E150R, E150V, E150W, E150Y, G151A, G151C, G151D, G151L,G151N, G151P, G151S, G151W, K152D, K152L, K152R, G164D, G164E, G164H,G164S, G164V, V167D, V167E, V167L, V167P, V167Q, V167R, V167W, H168N,L170A, L170D, L170E, L170F, L170H, L170K, L170M, L170N, L170P, L170Q,L170R, L170S, L170V, L170W, L170Y, D171A, D171C, D171E, D171K, D171L,D171M, D171Q, D171R, D171V, D171W, D171Y, I173C, D174H, D174M, D174N,D174V, D174W, F175Y, N177M, Q178*, Q178A, Q178K, Q178R, Q178W, I179T,S181C, S181D, S181F, S181G, S181N, S181P, S181Q, S181T, S181V, S181W,E185M, E185R, E185V, E185W, S186D, S186E, S186H, S186I, S186K, S186L,S186M, S186N, S186Q, S186R, S186V, S186W, K187C, K187D, K187G, K187R,K187S, K187V, K187W, Y188P, E189L, E189V, E189W, S199C, S199L, S199M,S199Y, E200D, E200F, E200K, E200L, E200M, E200N, E200R, E200W, A203C,A203D, A203E, A203G, A203L, A203M, A203P, A203R, A203S, A203T, A203V,A203W, N204L, N204M, N204V, N204W, N204Y, L205I, D207A, D207C, D207E,D207G, D207N, D207Q, D207S, D207V, D207W, S208A, S208C, S208D, S208G,S208L, S208Q, S208T, S208V, S208W, S210T, Q215R, Q215M, Q215L, Q215*,S217V, T218A, T218L, T218Q, T218R, T218V, G222D, E224A, E224P, N227A,N227Q, N227R, N227S, N227T, N227K, D230*, D230R, D230T, D230W, E232D,E232V, N233H, N233Q, N233R, N233W, W234R, G235W, G235A, G235E, G235F,G235H, G235I, G235L, G235M, G235N, G235P, G235S, G235V, S237C, S237G,S237M, S237N, S237W, S237Y, Y244C, Y244E, Y244M, L249H, L249K, L249Q,L249R, L249W, L249Y, S251L, S251N, S251R, S251W, N252P, N252C, G253D,G253W, F254I, F254L, F254M, F254Y, Q256E, Q256R, N260*, N260A, N260C,N260E, N260I, N260K, N260L, N260M, N260Q, N260R, N260V, N260W, N260Y,E261*, E261A, E261D, E261R, E261W, Q262*, Q262F, Q262H, Q262W, Q262Y,M263K, M263L, M263Q, D264*, D264C, D264E, Y265F, N267S, N267T, W268C,W268E, W268M, W268R, Y270F, A271D, A271G, H272D, H272I, H272M, H272P,H272V, H272W, N273W, K274R, K274A, K274H, F276A, F276C, F276K, F276N,F276G, F276L, F276M, F276P, F276S, F276V, F276W, I278A, I278K, I278N,I278Q, I278V, S279C, S279D, S279E, S279G, S279N, D280C, D280E, Y281*,Y281A, Y281C, Y281H, Y281K, Y281N, Y281P, Y281R, Y282E, Y282N, H283I,A284I, A284L, A284N, A284P, A284T, A284V, T287N, S288D, S288K, S288N,V290I, K291L, K291R, K291V, T296C, E300A, E300D, H301C, H301N, H301R,T303A, T303C, T303G, T303K, T303Q, T303R, T303W, D304C, D304M, L305M,L305N, S306C, K308A, K308D, K308G, K308I, K308L, K308Q, K308S, K308T,K308V, K308Y, K309A, K309C, K309D, K309H, K309L, K309M, K309N, K309Q,K309S, K309T, K309I, K312A, K312L, K312M, K312N, K312Q, K312S, K312W,E314I, E314L, E314V, L315I, L315V, R319A, and N323R, wherein the varianthas dispersin activity and wherein each position corresponds to theposition of the polypeptide shown in SEQ ID NO: 1 (numbering accordingto SEQ ID NO: 1).

In some preferred aspects the alteration is a substitution and thedispersin variant comprises one or more substitution(s) selected fromthe group consisting of: D2A, D2L, D2N, D2R, D2V, D2W, Q3F, Q3I, Q3L,Q3M, Q3P, Q3V, Q3Y, Q3T, S12A, H15F, H15Y, T17W, T17C, T17E, T17F, T17M,T17R, T17V, V18L, E19D, E19N, E19P, K22A, K22M, K22V, S23C, S23E, S23I,S23L, S23R, S23T, S23V, V25R, D26M, Y30D, Y30L, Y30M, Y30N, Y30R, Y30T,Y30V, G32L, G32M, G32R, N43H, N43L, N45D, N45L, N45V, A49W, A49Y, Y52M,G54L, G54M, G54N, S56T, S56W, S57W, E58N, N59A, N59C, N59D, N59E, N59F,N59M, N59R, N59V, N59W, T60V, N62C, N62D, N62H, N62Q, N62W, T63C, T63D,T63L, T63N, T63R, T63V, K67A, K67L, N68L, N68Q, L71H, L71N, L71R, L71V,L71W, S72C, S72D, S72E, S72F, S72G, S72I, S72M, S72N, S72R, S72T, S72Y,I74L, S77A, D79V, K80E, K80H, K80L, K80N, K80Q, K80V, K80W, D81A, D81G,D81L, D81R, D81S, D81T, D81V, D81W, I82V, L90F, E99Q, E99R, L100S,K103A, K103R, K104N, K104W, D105N, V106A, V106D, V106E, V106H, V106K,V106L, V106M, V106N, V106Q, V106W, V106Y, K107A, K107C, K107L, K107M,K107T, K107V, K107W, N110M, N110R, N110V, D111A, D111E, D111M, D111N,D111Q, D111R, D111V, D111W, V113T, T114C, T114S, Y116D, Y116N, Y116R,S117D, S117H, S117N, S117P, E118A, E118D, E118G, E118L, E119G, E119W,T120I, T120L, T120M, T120V, T120W, D122H, D122R, Y123W, Y124C, Y124I,Y124K, Y124L, Y124M, Y124Q, Y124R, Y124T, Y124V, Y124W, D125C, D125G,D125K, D125Q, D125R, N126V, R127D, R127H, R127K, R127L, R127M, R127Q,R127W, V128C, V128L, V128T, D131V, Q135A, Q135D, Q135E, Q135K, Q135M,Q135Y, D138K, D138L, D138M, D138Q, D138R, D138S, D138V, D138W, E139W,D142R, D142W, Y145H, Y145L, Y145N, Y145V, P147A, P147C, P147D, P147F,P147G, P147L, P147M, P147R, P147S, P147T, P147V, K148A, K148D, K148L,K148V, F149L, F149M, F149N, E150D, E150H, E150K, E150L, E150M, E150N,E150R, E150V, E150W, E150Y, G151A, G151C, G151D, G151L, G151N, G151P,G151S, G151W, K152D, K152L, K152R, G164D, G164E, G164H, G164S, G164V,V167D, V167E, V167L, V167P, V167Q, V167R, V167W, H168N, L170A, L170D,L170E, L170F, L170H, L170K, L170M, L170N, L170P, L170Q, L170R, L170S,L170V, L170W, L170Y, D171A, D171C, D171E, D171K, D171L, D171M, D171Q,D171R, D171V, D171W, D171Y, I173C, D174H, D174M, D174N, D174V, D174W,F175Y, N177M, Q178A, Q178K, Q178R, Q178W, I179T, S181C, S181D, S181F,S181G, S181N, S181P, S181Q, S181T, S181V, S181W, E185M, E185R, E185V,E185W, S186D, S186E, S186H, S186I, S186K, S186L, S186M, S186N, S186Q,S186R, S186V, S186W, K187C, K187D, K187G, K187R, K187S, K187V, K187W,Y188P, E189L, E189V, E189W, S199C, S199L, S199M, S199Y, E200D, E200F,E200K, E200L, E200M, E200N, E200R, E200W, A203C, A203D, A203E, A203G,A203L, A203M, A203P, A203R, A203S, A203T, A203V, A203W, N204L, N204M,N204V, N204W, N204Y, L205I, D207A, D207C, D207E, D207G, D207N, D207Q,D207S, D207V, D207W, S208A, S208C, S208D, S208G, S208L, S208Q, S208T,S208V, S208W, S210T, Q215R, Q215M, Q215L, S217V, T218A, T218L, T218Q,T218R, T218V, G222D, E224A, E224P, N227A, N227Q, N227R, N227S, N227T,N227K, D230R, D230T, D230W, E232D, E232V, N233H, N233Q, N233R, N233W,W234R, G235W, G235A, G235E, G235F, G235H, G235I, G235L, G235M, G235N,G235P, G235S, G235V, S237C, S237G, S237M, S237N, S237W, S237Y, Y244C,Y244E, Y244M, L249H, L249K, L249Q, L249R, L249W, L249Y, S251L, S251N,S251R, S251W, N252P, N252C, G253D, G253W, F254I, F254L, F254M, F254Y,Q256E, Q256R, N260A, N260C, N260E, N260I, N260K, N260L, N260M, N260Q,N260R, N260V, N260W, N260Y, E261A, E261D, E261R, E261W, Q262F, Q262H,Q262W, Q262Y, M263K, M263L, M263Q, D264C, D264E, Y265F, N267S, N267T,W268C, W268E, W268M, W268R, Y270F, A271D, A271G, H272D, H272I, H272M,H272P, H272V, H272W, N273W, K274R, K274A, K274H, F276A, F276C, F276K,F276N, F276G, F276L, F276M, F276P, F276S, F276V, F276W, I278A, I278K,I278N, I278Q, I278V, S279C, S279D, S279E, S279G, S279N, D280C, D280E,Y281A, Y281C, Y281H, Y281K, Y281N, Y281P, Y281R, Y282E, Y282N, H283I,A284I, A284L, A284N, A284P, A284T, A284V, T287N, S288D, S288K, S288N,V290I, K291L, K291R, K291V, T296C, E300A, E300D, H301C, H301N, H301R,T303A, T303C, T303G, T303K, T303Q, T303R, T303W, D304C, D304M, L305M,L305N, S306C, K308A, K308D, K308G, K308I, K308L, K308Q, K308S, K308T,K308V, K308Y, K309A, K309C, K309D, K309H, K309L, K309M, K309N, K309Q,K309S, K309T, K309I, K312A, K312L, K312M, K312N, K312Q, K312S, K312W,E314I, E314L, E314V, L315I, L315V, R319A, and N323R, wherein the varianthas dispersin activity and wherein each position corresponds to theposition of the polypeptide shown in SEQ ID NO: 1 (numbering accordingto SEQ ID NO: 1).

One aspect relates to a dispersin variant, which compared to a dispersinwith SEQ ID NO: 1, comprises one or more substitutions selected from thegroup consisting of: D2A, D2L, D2N, D2R, D2V, D2W, Q3F, Q3I, Q3L, Q3M,Q3P, Q3V, Q3Y, Q3T, S12A, H15F, H15Y, T17W, T17C, T17E, T17F, T17M,T17R, T17V, V18L, E19D, E19K, E19N, E19P, K22A, K22M, K22V, S23A, S23C,S23E, S23I, S23L, S23R, S23T, S23V, L24I, V25R, D26M, Y30A, Y30D, Y30L,Y30M, Y30N, Y30R, Y30T, Y30V, G32L, G32M, G32R, N34D, N43H, N43L, N45D,N45L, N45V, A49W, A49Y, Y52M, G54L, G54M, G54N, S56T, S56W, S57W, E58N,N59A, N59C, N59D, N59E, N59F, N59M, N59R, N59V, N59W, T60V, N62C, N62D,N62H, N62Q, N62W, T63C, T63D, T63L, T63N, T63R, T63V, K67A, K67L, N68L,N68Q, L71H, L71N, L71R, L71V, L71W, S72C, S72D, S72E, S72F, S72G, S72I,S72M, S72N, S72R, S72T, S72Y, I74L, S77A, D79V, K80E, K80H, K80L, K80N,K80Q, K80R, K80V, K80W, D81A, D81G, D81L, D81N, D81R, D81S, D81T, D81V,D81W, I82V, L90F, E99Q, E99R, L100S, K103A, K103R, K103V, K104N, K104W,D105N, V106A, V106D, V106E, V106H, V106K, V106L, V106M, V106N, V106Q,V106R, V106W, V106Y, K107A, K107C, K107L, K107M, K107T, K107V, K107W,N110M, N110R, N110V, D111A, D111E, D111M, D111N, D111Q, D111R, D111V,D111W, V113T, T114C, T114S, Y116D, Y116N, Y116R, S117D, S117H, S117N,S117P, E118A, E118D, E118G, E118L, E119G, E119W, T120I, T120L, T120M,T120V, T120W, D122H, D122R, Y123W, Y124C, Y124H, Y124I, Y124K, Y124L,Y124M, Y124N, Y124Q, Y124R, Y124T, Y124V, Y124W, D125C, D125G, D125H,D125K, D125Q, D125R, N126V, R127D, R127H, R127K, R127L, R127M, R127Q,R127W, V128A, V128C, V128D, V128L, V128T, D131V, Q135A, Q135D, Q135E,Q135K, Q135M, Q135Y, D138K, D138L, D138M, D138N, D138Q, D138R, D138S,D138V, D138W, E139W, V140I, D142R, D142W, Y145H, Y145L, Y145N, Y145V,P147A, P147C, P147D, P147F, P147G, P147L, P147M, P147R, P147S, P147T,P147V, K148A, K148D, K148L, K148V, F149L, F149M, F149N, E150A, E150D,E150H, E150K, E150L, E150M, E150N, E150R, E150V, E150W, E150Y, G151A,G151C, G151D, G151L, G151N, G151P, G151S, G151W, K152D, K152L, K152R,S163P, G164D, G164E, G164H, G164S, G164V, V167A, V167D, V167E, V167L,V167P, V167Q, V167R, V167W, H168N, L170A, L170D, L170E, L170F, L170H,L170K, L170M, L170N, L170P, L170Q, L170R, L170S, L170V, L170W, L170Y,D171A, D171C, D171E, D171K, D171L, D171M, D171Q, D171R, D171V, D171W,D171Y, I173C, D174H, D174M, D174N, D174V, D174W, F175Y, N177M, Q178A,Q178K, Q178R, Q178W, I179T, S181C, S181D, S181F, S181G, S181K, S181N,S181P, S181Q, S181T, S181V, S181W, E185A, E185M, E185R, E185V, E185W,S186D, S186E, S186H, S186I, S186K, S186L, S186M, S186N, S186Q, S186R,S186V, S186W, K187C, K187D, K187G, K187R, K187S, K187V, K187W, Y188P,E189L, E189V, E189W, S199C, S199L, S199M, S199Y, E200D, E200F, E200K,E200L, E200M, E200N, E200R, E200W, A203C, A203D, A203E, A203G, A203L,A203M, A203P, A203Q, A203R, A203S, A203T, A203V, A203W, N204L, N204M,N204V, N204W, N204Y, L205I, D207A, D207C, D207E, D207G, D207K, D207N,D207Q, D207R, D207S, D207V, D207W, S208A, S208C, S208D, S208G, S208L,S208Q, S208T, S208V, S208W, S210T, Q215K, Q215R, Q215M, Q215L, S217V,T218A, T218L, T218Q, T218R, T218V, S221N, G222D, E224A, E224P, S225G,N227A, N227Q, N227R, N227S, N227T, N227K, D230N, D230R, D230T, D230W,E232D, E232V, N233D, N233E, N233H, N233Q, N233R, N233W, W234R, G235A,G235W, G235E, G235F, G235H, G235I, G235L, G235M, G235N, G235P, G235S,G235V, S237C, S237G, S237M, S237N, S237W, S237Y, Y244C, Y244E, Y244M,L249H, L249K, L249Q, L249R, L249W, L249Y, S251A, S251L, S251N, S251R,S251W, N252P, N252C, G253D, G253W, F254I, F254L, F254M, F254Y, Q256D,Q256E, Q256R, N260A, N260C, N260E, N260I, N260K, N260L, N260M, N260Q,N260R, N260T, N260V, N260W, N260Y, E261A, E261D, E261R, E261W, Q262F,Q262H, Q262W, Q262Y, M263K, M263L, M263Q, D264C, D264E, D264N, Y265F,N267S, N267T, W268C, W268E, W268M, W268R, Y270F, A271D, A271G, H272D,H272I, H272M, H272P, H272V, H272W, N273W, K274R, K274A, K274H, F276A,F276C, F276K, F276N, F276G, F276L, F276M, F276P, F276S, F276V, F276W,I278A, I278K, I278N, I278Q, I278V, S279C, S279D, S279E, S279G, S279N,D280C, D280E, Y281A, Y281C, Y281H, Y281K, Y281N, Y281P, Y281R, Y282E,Y282N, H283I, A284I, A284L, A284N, A284P, A284T, A284V, T287N, S288P,S288D, S288K, S288N, V290I, K291L, K291R, K291V, T296C, E300A, E300D,H301C, H301N, H301R, T303A, T303C, T303G, T303K, T303Q, T303R, T303W,D304C, D304M, L305M, L305N, S306C, K308A, K308D, K308E, K308G, K308I,K308L, K308Q, K308S, K308T, K308V, K308Y, K309A, K309C, K309D, K309E,K309G, K309H, K309L, K309M, K309N, K309Q, K309S, K309T, K309I, K312A,K312E, K312L, K312M, K312N, K312Q, K312S, K312W, E314I, E314L, E314V,L315I, L315V, R319A, Y321F and N323R, wherein the positions correspondto the positions of SEQ ID NO: 1 (numbering according to SEQ ID NO: 1)wherein the variant has a sequence identity to the polypeptide shown inSEQ ID NO: of at least at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 95%, suchas at least 96%, at least 97%, at least 98%, or at least 99%, but lessthan 100%, sequence identity to SEQ ID NO: 1, and wherein the varianthas dispersin, preferably beta-1,6 N-acetylglucosaminidase activity.

One aspect relates to a dispersin variant, which compared to a dispersinwith SEQ ID NO: 1, comprises one or more substitutions selected from thegroup consisting of: D2A, D2L, D2N, D2R, D2V, D2W, Q3F, Q3I, Q3L, Q3M,Q3P, Q3V, Q3Y, Q3T, S12A, H15F, H15Y, T17W, T17C, T17E, T17F, T17M,T17R, T17V, V18L, E19D, E19N, E19P, K22A, K22M, K22V, S23C, S23E, S23I,S23L, S23R, S23T, S23V, V25R, D26M, Y30D, Y30L, Y30M, Y30N, Y30R, Y30T,Y30V, G32L, G32M, G32R, N43H, N43L, N45D, N45L, N45V, A49W, A49Y, Y52M,G54L, G54M, G54N, S56T, S56W, S57W, E58N, N59A, N59C, N59D, N59E, N59F,N59M, N59R, N59V, N59W, T60V, N62C, N62D, N62H, N62Q, N62W, T63C, T63D,T63L, T63N, T63R, T63V, K67A, K67L, N68L, N68Q, L71H, L71N, L71R, L71V,L71W, S72C, S72D, S72E, S72F, S72G, S72I, S72M, S72N, S72R, S72T, S72Y,I74L, S77A, D79V, K80E, K80H, K80L, K80N, K80Q, K80V, K80W, D81A, D81G,D81L, D81R, D81S, D81T, D81V, D81W, I82V, L90F, E99Q, E99R, L100S,K103A, K103R, K104N, K104W, D105N, V106A, V106D, V106E, V106H, V106K,V106L, V106M, V106N, V106Q, V106W, V106Y, K107A, K107C, K107L, K107M,K107T, K107V, K107W, N110M, N110R, N110V, D111A, D111E, D111M, D111N,D111Q, D111R, D111V, D111W, V113T, T114C, T114S, Y116D, Y116N, Y116R,S117D, S117H, S117N, S117P, E118A, E118D, E118G, E118L, E119G, E119W,T120I, T120L, T120M, T120V, T120W, D122H, D122R, Y123W, Y124C, Y124I,Y124K, Y124L, Y124M, Y124Q, Y124R, Y124T, Y124V, Y124W, D125C, D125G,D125K, D125Q, D125R, N126V, R127D, R127H, R127K, R127L, R127M, R127Q,R127W, V128C, V128L, V128T, D131V, Q135A, Q135D, Q135E, Q135K, Q135M,Q135Y, D138K, D138L, D138M, D138Q, D138R, D138S, D138V, D138W, E139W,D142R, D142W, Y145H, Y145L, Y145N, Y145V, P147A, P147C, P147D, P147F,P147G, P147L, P147M, P147R, P147S, P147T, P147V, K148A, K148D, K148L,K148V, F149L, F149M, F149N, E150D, E150H, E150K, E150L, E150M, E150N,E150R, E150V, E150W, E150Y, G151A, G151C, G151D, G151L, G151N, G151P,G151S, G151W, K152D, K152L, K152R, G164D, G164E, G164H, G164S, G164V,V167D, V167E, V167L, V167P, V167Q, V167R, V167W, H168N, L170A, L170D,L170E, L170F, L170H, L170K, L170M, L170N, L170P, L170Q, L170R, L170S,L170V, L170W, L170Y, D171A, D171C, D171E, D171K, D171L, D171M, D171Q,D171R, D171V, D171W, D171Y, I173C, D174H, D174M, D174N, D174V, D174W,F175Y, N177M, Q178A, Q178K, Q178R, Q178W, I179T, S181C, S181D, S181F,S181G, S181N, S181P, S181Q, S181T, S181V, S181W, E185M, E185R, E185V,E185W, S186D, S186E, S186H, S186I, S186K, S186L, S186M, S186N, S186Q,S186R, S186V, S186W, K187C, K187D, K187G, K187R, K187S, K187V, K187W,Y188P, E189L, E189V, E189W, S199C, S199L, S199M, S199Y, E200D, E200F,E200K, E200L, E200M, E200N, E200R, E200W, A203C, A203D, A203E, A203G,A203L, A203M, A203P, A203R, A2035, A203T, A203V, A203W, N204L, N204M,N204V, N204W, N204Y, L205I, D207A, D207C, D207E, D207G, D207N, D207Q,D207S, D207V, D207W, S208A, S208C, S 208D, S208G, S208L, S208Q, S208T,S208V, S208W, S210T, Q215R, Q215M, Q215L, S217V, T218A, T218L, T218Q,T218R, T218V, G222D, E224A, E224P, N227A, N227Q, N227R, N227S, N227T,N227K, D230R, D230T, D230W, E232D, E232V, N233H, N233Q, N233R, N233W,W234R, G235W, G235A, G235E, G235F, G235H, G235I, G235L, G235M, G235N,G235P, G235S, G235V, S237C, S237G, S237M, S237N, S237W, S237Y, Y244C,Y244E, Y244M, L249H, L249K, L249Q, L249R, L249W, L249Y, S251L, S251N,S251R, S251W, N252P, N252C, G253D, G253W, F254I, F254L, F254M, F254Y,Q256E, Q256R, N260A, N260C, N260E, N260I, N260K, N260L, N260M, N260Q,N260R, N260V, N260W, N260Y, E261A, E261D, E261R, E261W, Q262F, Q262H,Q262W, Q262Y, M263K, M263L, M263Q, D264C, D264E, Y265F, N267S, N267T,W268C, W268E, W268M, W268R, Y270F, A271D, A271G, H272D, H272I, H272M,H272P, H272V, H272W, N273W, K274R, K274A, K274H, F276A, F276C, F276K,F276N, F276G, F276L, F276M, F276P, F276S, F276V, F276W, I278A, I278K,I278N, I278Q, I278V, S279C, S279D, S279E, S279G, S279N, D280C, D280E,Y281A, Y281C, Y281H, Y281K, Y281N, Y281P, Y281R, Y282E, Y282N, H283I,A284I, A284L, A284N, A284P, A284T, A284V, T287N, S288D, S288K, S288N,V290I, K291L, K291R, K291V, T296C, E300A, E300D, H301C, H301N, H301R,T303A, T303C, T303G, T303K, T303Q, T303R, T303W, D304C, D304M, L305M,L305N, S306C, K308A, K308D, K308G, K308I, K308L, K308Q, K308S, K308T,K308V, K308Y, K309A, K309C, K309D, K309H, K309L, K309M, K309N, K309Q,K309S, K309T, K309I, K312A, K312L, K312M, K312N, K312Q, K312S, K312W,E314I, E314L, E314V, L315I, L315V, R319A, and N323R, wherein thepositions correspond to the positions of SEQ ID NO: 1 (numberingaccording to SEQ ID NO: 1) wherein the variant has a sequence identityto the polypeptide shown in SEQ ID NO: 1 of at least at least 60%, atleast 65%, at least 70%, at least 75%, at least 80%, at least 85%, atleast 90%, at least 95%, such as at least 96%, at least 97%, at least98%, or at least 99%, but less than 100%, sequence identity to SEQ IDNO: 1, and wherein the variant has dispersin, preferably beta-1,6N-acetylglucosaminidase activity.

In some preferred aspects the alteration is a substitution and thedispersin variant comprises one or more substitutions selected from thegroup consisting of: D2V, Q3F, Q3I, S12A, H15Y, T17W, T17E, V18L, K22M,S23I, S23C, S23T, S23L, S23V, S23E, V25R, Y30L, A49W, A49Y, S56T, N59D,N59C, N59E, N59R, N59F, N59W, N59V, N62C, N62D, T63C, N68Q, S72D, S72E,I74L, S77A, I82V, L90F, E99Q, L100S, T114S, T114C, Y123W, Y124I, Y124M,Y124R, Y124V, Y124Q, Y124T, Y124K, D125R, D125C, D125G, D125K, D125Q,Q135M, D138R, D138K, D138Q, L170D, L170K, L170S, L170H, D171E, D171K,D171Y, D171M, D171Q, D171L, I173C, D174W, D174H, D174M, D174N, F175Y,Q178K, I179T, S181T, S181F, S181Q, S181G, S181N, S181C, E185R, E185M,E185V, S186K, S186M, S186R, S186H, K187G, Y188P, E189V, S199C, S199L,A203G, A203E, N227T, N227K, E232D, G235W, S237W, Y244M, Y244C, N252P,N252C, Q256E, N260Q, Q262H, M263Q, D264E, Y265F, N267T, N267S, Y270F,H272M, H272P, H272I, H272V, N273W, K274H, F276A, F276N, F276K, F276C,I278V, S279N, S279D, S279G, D280E, D280C, Y281P, Y282N, H283I, A284T,A284L, A284I, S288K, V290I, K291R, T296C, T303Q, D304M, D304C, L305M,L305N, S306C, K308D, K308A, K308V, K308Q, K308S, K308Y, K308G, K308L,K308T, K308I, K309C, K309L, K309D, K309Q, K309N, K309T, K309A, K309S,K309M, K309H, K312A, K312Q, K312S, K312W, K312L, K312N, E314L, E314V,E314I, L315I, R319A and N323R, wherein the variant has dispersinactivity, preferably beta-1,6 N-acetylglucosaminidase activity andwherein each position corresponds to the position of the polypeptideshown in SEQ ID NO: 1 (numbering according to SEQ ID NO: 1).

In some aspects the variant has a sequence identity of at least 60%, atleast 65%, at least 70%, at least 75%, at least 80%, at least 85%, atleast 90%, at least 95%, such as at least 96%, at least 97%, at least98%, or at least 99%, but less than 100%, to the amino acid sequence ofthe mature parent dispersin e.g. the dispersin shown in SEQ ID NO: 1.

In some aspects, the number of substitutions in the variants of thepresent invention is 2-20, e.g., 2-10 and 2-5, such as 2, 3, 4, 5, 6, 7,8, 9 ,10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 substitutions. In oneaspect the total number of substitutions compared to SEQ ID NO: 1 is3-20, e.g. at least 5 to 20, at least 10 to 20 or such as at least 15 to20 substitutions compared to the dispersin comprising the amino acidsequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 2 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 2 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Val, Leu, Ala, Trp, Arg or Asn. In another aspect,the dispersin variant comprises or consists of a substitution selectedfrom the group consisting of D2V, D2L, D2A, D2W, D2R and D2N, whereinthe position corresponds to the positions of SEQ ID NO: 1 and whereinthe variant has at least 60%, at least 65%, at least 70%, at least 75%,at least 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 3 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 3 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Phe, Leu, Val, Tyr, Met, Ile, Pro or Thr. Inanother aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of Q3F, Q3L, Q3V, Q3Y,Q3M, Q3I, Q3P and Q3T, wherein the position corresponds to the positionsof SEQ ID NO: 1 and wherein the variant has at least at least 60%, atleast 65%, at least 70%, at least 75%, at least 80%, at least 85%, atleast 90%, at least 95%, such as at least 96%, at least 97%, at least98%, or at least 99%, but less than 100%, sequence identity to the aminoacid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 12 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 12 of SEQ ID NO: 1 is substituted with Ala, Ar , Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Ala. In another aspect, the dispersin variantcomprises or consists of the substitution S12A, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 15 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 15 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Tyr or Phe. In another aspect, the dispersinvariant comprises or consists of the substitution H15Y or H15F, whereinthe position corresponds to the positions of SEQ ID NO: 1 and whereinthe variant has at least at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 95%, suchas at least 96%, at least 97%, at least 98%, or at least 99%, but lessthan 100%, sequence identity to the amino acid sequence shown in SEQ IDNO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 17 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 17 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Trp, Val, Arg, Met, Phe, Cys or Glu. In anotheraspect, the dispersin variant comprises or consists of a substitutionselected from the group consisting of T17W, T17V, T17R, T17M, T17F, T17Cand T17E, wherein the position corresponds to the positions of SEQ IDNO: 1 and wherein the variant has at least at least 60%, at least 65%,at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 18 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 18 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Leu. In another aspect, the dispersin variantcomprises or consists of the substitution V18L, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 19 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 19 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Pro, Asn, Lys or Asp. In another aspect, thedispersin variant comprises or consists of a substitution selected fromthe group consisting of E19P, E19N, E19K and E19D, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 22 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 22 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Ala, Val or Met. In another aspect, the dispersinvariant comprises or consists of a substitution selected from the groupconsisting of K22A, K22V and K22M, wherein the position corresponds tothe positions of SEQ ID NO: 1 and wherein the variant has at least atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100%, sequenceidentity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 23 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 23 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Ala, Cys, Glu, Ile, Leu, Arg, Thr or Val. Inanother aspect, the dispersin variant comprises or consists of thesubstitution selected from the group consisting of S23A, S23C, S23E,S23I, S23L, S23R, S23T and S23V, wherein the position corresponds to thepositions of SEQ ID NO: 1 and wherein the variant has at least at least60%, at least 65%, at least 70%, at least 75%, at least 80%, at least85%, at least 90%, at least 95%, such as at least 96%, at least 97%, atleast 98%, or at least 99%, but less than 100%, sequence identity to theamino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 24 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 24 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Ile. In another aspect, the dispersin variantcomprises or consists of the substitution L24I, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 25 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 25 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Arg. In another aspect, the dispersin variantcomprises or consists of the substitution V25R, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 26 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 26 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Met. In another aspect, the dispersin variantcomprises or consists of the substitution D26M, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of adeletion of the amino acid at a position corresponding to position 30 ofSEQ ID NO: 1. In another aspect, the dispersin variant comprises orconsists of a substitution at a position corresponding to position 30 ofSEQ ID NO: 1. In another aspect, the amino acid at a positioncorresponding to position 30 of SEQ ID NO: 1 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val, preferably with Arg, Val, Asp, Thr, Asn,Met, Ala or Leu. In another aspect, the dispersin variant comprises orconsists of a substitution selected from the group consisting of Y30R,Y30V, Y30D, Y30T, Y30N, Y30M, Y30A and Y30L, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 32 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 32 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Arg, Met or Leu. In another aspect, the dispersinvariant comprises or consists of a substitution selected from the groupconsisting of G32R, G32M and G32L, wherein the position corresponds tothe positions of SEQ ID NO: 1 and wherein the variant has at least atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100%, sequenceidentity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 34 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 34 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Asp. In another aspect, the dispersin variantcomprises or consists of the substitution N34D, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of adeletion of the amino acid at a position corresponding to position 43 ofSEQ ID NO: 1. In another aspect, the dispersin variant comprises orconsists of a substitution at a position corresponding to position 43 ofSEQ ID NO: 1. In another aspect, the amino acid at a positioncorresponding to position 43 of SEQ ID NO: 1 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val, preferably with Leu or His. In anotheraspect, the dispersin variant comprises or consists of the substitutionN43L or N43H, wherein the position corresponds to the positions of SEQID NO: 1 and wherein the variant has at least at least 60%, at least65%, at least 70%, at least 75%, at least 80%, at least 85%, at least90%, at least 95%, such as at least 96%, at least 97%, at least 98%, orat least 99%, but less than 100%, sequence identity to the amino acidsequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of adeletion of the amino acid at a position corresponding to position 44 ofSEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 45 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 45 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Asp, Leu or Val. In another aspect, the dispersinvariant comprises or consists of a substitution selected from the groupconsisting of N45D, N45L and N45V, wherein the position corresponds tothe positions of SEQ ID NO: 1 and wherein the variant has at least atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100%, sequenceidentity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 49 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 49 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Trp or Tyr. In another aspect, the dispersinvariant comprises or consists of the substitution A49W, A49Y, whereinthe position corresponds to the positions of SEQ ID NO: 1 and whereinthe variant has at least at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 95%, suchas at least 96%, at least 97%, at least 98%, or at least 99%, but lessthan 100%, sequence identity to the amino acid sequence shown in SEQ IDNO: 1.

In another aspect, the dispersin variant comprises or consists of adeletion of the amino acid at a position corresponding to position 52 ofSEQ ID NO: 1. In another aspect, the dispersin variant comprises orconsists of a substitution at a position corresponding to position 52 ofSEQ ID NO: 1. In another aspect, the amino acid at a positioncorresponding to position 52 of SEQ ID NO: 1 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val, preferably with Met. In another aspect, thedispersin variant comprises or consists of the substitution Y52M,wherein the position corresponds to the positions of SEQ ID NO: 1 andwherein the variant has at least at least 60%, at least 65%, at least70%, at least 75%, at least 80%, at least 85%, at least 90%, at least95%, such as at least 96%, at least 97%, at least 98%, or at least 99%,but less than 100%, sequence identity to the amino acid sequence shownin SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 54 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 54 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Asn, Leu or Met. In another aspect, the dispersinvariant comprises or consists of a substitution selected from the groupconsisting of G54N, G54L and G54M, wherein the position corresponds tothe positions of SEQ ID NO: 1 and wherein the variant has at least atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100%, sequenceidentity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 56 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 56 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Trp or Thr. In another aspect, the dispersinvariant comprises or consists of the substitution S56W or S56T, whereinthe position corresponds to the positions of SEQ ID NO: 1 and whereinthe variant has at least at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 95%, suchas at least 96%, at least 97%, at least 98%, or at least 99%, but lessthan 100%, sequence identity to the amino acid sequence shown in SEQ IDNO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 57 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 57 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Trp. In another aspect, the dispersin variantcomprises or consists of the substitution S57W, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 58 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 58 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Asn. In another aspect, the dispersin variantcomprises or consists of the substitution E58N, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 59 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 59 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Ala, Cys, Asp, Glu, Phe, Met, Arg, Val or Trp. Inanother aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of N59A, N59C, N59D,N59E, N59F, N59M, N59R, N59V, N59W, wherein the position corresponds tothe positions of SEQ ID NO: 1 and wherein the variant has at least atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100%, sequenceidentity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 60 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 60 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Val. In another aspect, the dispersin variantcomprises or consists of the substitution T60V, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 62 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 62 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Trp, Gln, His, Cys or Asp. In another aspect, thedispersin variant comprises or consists of a substitution selected fromthe group consisting of N62W, N62Q, N62H, N62C and N62D, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 63 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 63 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Val, Leu, Asn, Arg, Asp or Cys. In another aspect,the dispersin variant comprises or consists of a substitution selectedfrom the group consisting of T63V, T63L, T63N, T63R, T63D and T63C,wherein the position corresponds to the positions of SEQ ID NO: 1 andwherein the variant has at least at least 60%, at least 65%, at least70%, at least 75%, at least 80%, at least 85%, at least 90%, at least95%, such as at least 96%, at least 97%, at least 98%, or at least 99%,but less than 100%, sequence identity to the amino acid sequence shownin SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 67 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 67 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Leu or Ala. In another aspect, the dispersinvariant comprises or consists of the substitution K67L or K67A, whereinthe position corresponds to the positions of SEQ ID NO: 1 and whereinthe variant has at least at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 95%, suchas at least 96%, at least 97%, at least 98%, or at least 99%, but lessthan 100%, sequence identity to the amino acid sequence shown in SEQ IDNO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 68 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 68 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Leu or Gln. In another aspect, the dispersinvariant comprises or consists of the substitution N68L or N68Q, whereinthe position corresponds to the positions of SEQ ID NO: 1 and whereinthe variant has at least at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 95%, suchas at least 96%, at least 97%, at least 98%, or at least 99%, but lessthan 100%, sequence identity to the amino acid sequence shown in SEQ IDNO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 71 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 71 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Arg, His, Asn, Trp, or Val. In another aspect, thedispersin variant comprises or consists of a substitution selected fromthe group consisting of L71R, L71H, L71N, L71W and L71V, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of adeletion of the amino acid at a position corresponding to position 72 ofSEQ ID NO: 1. In another aspect, the dispersin variant comprises orconsists of a substitution at a position corresponding to position 72 ofSEQ ID NO: 1. In another aspect, the amino acid at a positioncorresponding to position 72 of SEQ ID NO: 1 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val, preferably with Thr, Gly, Cys, Met, Phe,Asn, Tyr, Arg, Ile, Asp or Glu. In another aspect, the dispersin variantcomprises or consists of a substitution selected from the groupconsisting of S72T, S72G, S72C, S72M, S72F, S72N, S72Y, S72R, S72I, S72Dand S72E, wherein the position corresponds to the positions of SEQ IDNO: 1 and wherein the variant has at least at least 60%, at least 65%,at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 74 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 74 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Leu. In another aspect, the dispersin variantcomprises or consists of the substitution I74L, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 77 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 77 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Ala. In another aspect, the dispersin variantcomprises or consists of the substitution S77A, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 79 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 79 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Val. In another aspect, the dispersin variantcomprises or consists of the substitution D79V, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of adeletion of the amino acid at a position corresponding to position 80 ofSEQ ID NO: 1. In another aspect, the dispersin variant comprises orconsists of a substitution at a position corresponding to position 80 ofSEQ ID NO: 1. In another aspect, the amino acid at a positioncorresponding to position 80 of SEQ ID NO: 1 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val, preferably with Asn, Arg, Trp, Leu, Val, Glnor His. In another aspect, the dispersin variant comprises or consistsof a substitution selected from the group consisting of K80N, K80R,K80W, K80E, K80L, K80V, K80Q and K80H, wherein the position correspondsto the positions of SEQ ID NO: 1 and wherein the variant has at least atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100%, sequenceidentity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 81 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 81 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Gly, Ala, Trp, Leu, Trp, Thr, Val, Arg, Ser or Asn.In another aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of D81G, D81A, D81L,D81W, D81T, D81V, D81R, D81S and D81N, wherein the position correspondsto the positions of SEQ ID NO: 1 and wherein the variant has at least atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100%, sequenceidentity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 82 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 82 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Val. In another aspect, the dispersin variantcomprises or consists of the substitution I82V, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 90 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 90 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Phe. In another aspect, the dispersin variantcomprises or consists of the substitution L90F, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 99 of SEQ ID NO: 1.In another aspect, the amino acid at a position corresponding toposition 99 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp, Cys,Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, orVal, preferably with Gln or Arg. In another aspect, the dispersinvariant comprises or consists of the substitution E99Q or E99R, whereinthe position corresponds to the positions of SEQ ID NO: 1 and whereinthe variant has at least at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 95%, suchas at least 96%, at least 97%, at least 98%, or at least 99%, but lessthan 100%, sequence identity to the amino acid sequence shown in SEQ IDNO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 100 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 100 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Ser. In another aspect, the dispersinvariant comprises or consists of the substitution L100S, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 103 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 103 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Arg, Val or Ala. In another aspect, thedispersin variant comprises or consists of a substitution selected fromthe group consisting of K103R, K103V and K103A, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 104 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 104 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asn or Trp. In another aspect, thedispersin variant comprises or consists of the substitution K104N orK104W, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 105 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 105 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asn. In another aspect, the dispersinvariant comprises or consists of the substitution D105N, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 106 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 106 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Tyr, Leu, His, Glu, Trp, Asn, Arg, Met,Asp, Lys, Ala or Gln. In another aspect, the dispersin variant comprisesor consists of a substitution selected from the group consisting ofV106Y, V106L, V106H, V106E, V106W, V106N, V106R, V106M, V106D, V106K,V106A and V106Q, wherein the position corresponds to the positions ofSEQ ID NO: 1 and wherein the variant has at least at least 60%, at least65%, at least 70%, at least 75%, at least 80%, at least 85%, at least90%, at least 95%, such as at least 96%, at least 97%, at least 98%, orat least 99%, but less than 100%, sequence identity to the amino acidsequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 107 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 107 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Met, Trp, Leu, Cys, Val, Ala or Thr. Inanother aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of K107M, K107W, K107L,K107C, K107V, K107A and K107T, wherein the position corresponds to thepositions of SEQ ID NO: 1 and wherein the variant has at least at least60%, at least 65%, at least 70%, at least 75%, at least 80%, at least85%, at least 90%, at least 95%, such as at least 96%, at least 97%, atleast 98%, or at least 99%, but less than 100%, sequence identity to theamino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 110 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 110 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Arg, Val or Met. In another aspect, thedispersin variant comprises or consists of a substitution selected fromthe group consisting of: N110R, N110V and N110M, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 111 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 111 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Val, Ala, Arg, Asn, Trp, Met, Gln or Glu.In another aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of: D111V, D111A, D111R,D111N, D111W, D111M, D111Q and D111E, wherein the position correspondsto the positions of SEQ ID NO: 1 and wherein the variant has at least atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100%, sequenceidentity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 113 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 113 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Thr. In another aspect, the dispersinvariant comprises or consists of the substitution V113T, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 114 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 114 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Ser or Cys. In another aspect, thedispersin variant comprises or consists of the substitution T114S orT114C, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 116 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 116 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asp, Asn or Arg. In another aspect, thedispersin variant comprises or consists of the substitution Y116D, Y116Nor Y116R, wherein the position corresponds to the positions of SEQ IDNO: 1 and wherein the variant has at least at least 60%, at least 65%,at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of adeletion of the amino acid at a position corresponding to position 117of SEQ ID NO: 1. In another aspect, the dispersin variant comprises orconsists of a substitution at a position corresponding to position 117of SEQ ID NO: 1. In another aspect, the amino acid at a positioncorresponding to position 117 of SEQ ID NO: 1 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val, preferably with Asp, His, Asn or Pro. Inanother aspect, the dispersin variant comprises or consists of thesubstitution S117D, S117H, S117N or S117P, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of adeletion of the amino acid at a position corresponding to position 118of SEQ ID NO: 1. In another aspect, the dispersin variant comprises orconsists of a substitution at a position corresponding to position 118of SEQ ID NO: 1. In another aspect, the amino acid at a positioncorresponding to position 118 of SEQ ID NO: 1 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val, preferably with Ala, Asp, Gly or Leu. Inanother aspect, the dispersin variant comprises or consists of thesubstitution E118A, E118D, E118G or E118L, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 119 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 119 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Gly or Trp. In another aspect, thedispersin variant comprises or consists of the substitution E119G orE119W, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 120 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 120 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Ile, Leu, Met, Val or Trp. In anotheraspect, the dispersin variant comprises or consists of the substitutionT120I, T120L, T120M, T120V or T120W, wherein the position corresponds tothe positions of SEQ ID NO: 1 and wherein the variant has at least atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100%, sequenceidentity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of adeletion of the amino acid at a position corresponding to position 122of SEQ ID NO: 1. In another aspect, the dispersin variant comprises orconsists of a substitution at a position corresponding to position 122of SEQ ID NO: 1. In another aspect, the amino acid at a positioncorresponding to position 122 of SEQ ID NO: 1 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val, preferably with Arg or His. In anotheraspect, the dispersin variant comprises or consists of the substitutionD122R or D122H, wherein the position corresponds to the positions of SEQID NO: 1 and wherein the variant has at least at least 60%, at least65%, at least 70%, at least 75%, at least 80%, at least 85%, at least90%, at least 95%, such as at least 96%, at least 97%, at least 98%, orat least 99%, but less than 100%, sequence identity to the amino acidsequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 123 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 123 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Trp. In another aspect, the dispersinvariant comprises or consists of the substitution Y123W, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 124 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 124 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Ile, Met, His, Arg, Val, Gln, Thr, Lys,Leu, Asn, Trp or Cys. In another aspect, the dispersin variant comprisesor consists of a substitution selected from the group consisting of:Y124I, Y124M, Y124H, Y124R, Y124V, Y124Q, Y124T, Y124K, Y124L, Y124N,Y124W and Y124C, wherein the position corresponds to the positions ofSEQ ID NO: 1 and wherein the variant has at least at least 60%, at least65%, at least 70%, at least 75%, at least 80%, at least 85%, at least90%, at least 95%, such as at least 96%, at least 97%, at least 98%, orat least 99%, but less than 100%, sequence identity to the amino acidsequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 125 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 125 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Arg, Cys, Gly, His, Lys or Gln. In anotheraspect, the dispersin variant comprises or consists of a substitutionselected from the group consisting of: D125R, D125C, D125G, D125H, D125Kand D125Q, wherein the position corresponds to the positions of SEQ IDNO: 1 and wherein the variant has at least at least 60%, at least 65%,at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 126 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 126 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Val. In another aspect, the dispersinvariant comprises or consists of the substitution N126V, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 127 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 127 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Leu, Trp, Gln, Met, Asp, His or Lys. Inanother aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of: R127L, R127W, R127Q,R127M, R127D, R127H and R127K, wherein the position corresponds to thepositions of SEQ ID NO: 1 and wherein the variant has at least at least60%, at least 65%, at least 70%, at least 75%, at least 80%, at least85%, at least 90%, at least 95%, such as at least 96%, at least 97%, atleast 98%, or at least 99%, but less than 100%, sequence identity to theamino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 128 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 128 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Leu, Asp, Ala, Cys, or Thr. In anotheraspect, the dispersin variant comprises or consists of a substitutionselected from the group consisting of: V128L, V128D, V128A, V128C andV128T, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 131 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 131 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Val. In another aspect, the dispersinvariant comprises or consists of the substitution D131V, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of adeletion of the amino acid at a position corresponding to position 135of SEQ ID NO: 1. In another aspect, the dispersin variant comprises orconsists of a substitution at a position corresponding to position 135of SEQ ID NO: 1. In another aspect, the amino acid at a positioncorresponding to position 135 of SEQ ID NO: 1 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val, preferably with Met, Tyr, Ala, Asp, Glu orLys. In another aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of: Q135M, Q135Y, Q135A,Q135D, Q135E and Q135K, wherein the position corresponds to thepositions of SEQ ID NO: 1 and wherein the variant has at least at least60%, at least 65%, at least 70%, at least 75%, at least 80%, at least85%, at least 90%, at least 95%, such as at least 96%, at least 97%, atleast 98%, or at least 99%, but less than 100%, sequence identity to theamino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 138 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 138 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Met, Val, Leu, Trp, Ser, Arg, Lys, Asn orGln. In another aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of: D138M, D138V, D138L,D138W, D138S, D138R, D138K, D138N and D138Q, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 139 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 139 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Trp. In another aspect, the dispersinvariant comprises or consists of the substitution E139W, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 140 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 140 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Ile. In another aspect, the dispersinvariant comprises or consists of the substitution V140I, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 142 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 142 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Trp or Arg. In another aspect, thedispersin variant comprises or consists of the substitution D142W orD142R, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1

In another aspect, the dispersin variant comprises or consists of adeletion of the amino acid at a position corresponding to position 145of SEQ ID NO: 1. In another aspect, the dispersin variant comprises orconsists of a substitution at a position corresponding to position 145of SEQ ID NO: 1. In another aspect, the amino acid at a positioncorresponding to position 145 of SEQ ID NO: 1 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val, preferably with Val, Asn, His or Leu. Inanother aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of: Y145V, Y145N, Y145Hand Y145L, wherein the position corresponds to the positions of SEQ IDNO: 1 and wherein the variant has at least at least 60%, at least 65%,at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 147 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 147 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Leu, Asp, Ala, Val, Gly, Arg, Ser, Met,Thr, Cys or Phe. In another aspect, the dispersin variant comprises orconsists of a substitution selected from the group consisting of: P147L,P147D, P147A, P147V, P147G, P147R, P147S, P147M, P147T, P147C and P147F,wherein the position corresponds to the positions of SEQ ID NO: 1 andwherein the variant has at least at least 60%, at least 65%, at least70%, at least 75%, at least 80%, at least 85%, at least 90%, at least95%, such as at least 96%, at least 97%, at least 98%, or at least 99%,but less than 100%, sequence identity to the amino acid sequence shownin SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 148 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 148 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Leu, Asp, Ala or Val. In another aspect,the dispersin variant comprises or consists of a substitution selectedfrom the group consisting of: K148L, K148D, K148A and K148V, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 149 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 149 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asn, Met or Leu. In another aspect, thedispersin variant comprises or consists of a substitution selected fromthe group consisting of: F149N, F149M and F149L, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 150 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 150 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asn, Arg, Asp, Lys, Mat, His, Trp, Val,Gly, Leu or Ala. In another aspect, the dispersin variant comprises orconsists of a substitution selected from the group consisting of: E150N,E150R, E150D, E150K, E150M, E150H, E150W, E150V, E150Y, E150L and E150A,wherein the position corresponds to the positions of SEQ ID NO: 1 andwherein the variant has at least at least 60%, at least 65%, at least70%, at least 75%, at least 80%, at least 85%, at least 90%, at least95%, such as at least 96%, at least 97%, at least 98%, or at least 99%,but less than 100%, sequence identity to the amino acid sequence shownin SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 151 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 151 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Ala, Pro, Trp, Asp, Leu, Ser, Asn or Cys.In another aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of: G151A, G151P, G151W,G151D, G151L, G151S, G151N and G151C, wherein the position correspondsto the positions of SEQ ID NO: 1 and wherein the variant has at least atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100%, sequenceidentity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 152 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 152 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asp, Leu or Arg. In another aspect, thedispersin variant comprises or consists of a substitution selected fromthe group consisting of: K152D, K152L and K152R, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 163 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 163 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Pro. In another aspect, the dispersinvariant comprises or consists of the substitution S163P, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO: 1

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 164 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 164 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asp, His, Ser, Glu or Val. In anotheraspect, the dispersin variant comprises or consists of a substitutionselected from the group consisting of: G164D, G164H, G164S, G164E andG164V, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 167 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 167 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Gln, Ala, Trp, Pro, Asp, Leu, Arg or Glu.In another aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of: V167Q, V167A, V167W,V167P, V167D, V167L, V167R and V167E, wherein the position correspondsto the positions of SEQ ID NO: 1 and wherein the variant has at least atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100%, sequenceidentity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 168 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 168 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asn. In another aspect, the dispersinvariant comprises or consists of the substitution H168N, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO: 1

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 170 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 170 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asp, Lys, Ser, His, Asn, Arg, Glu, Phe,Met, Ala, Trp, Tyr, Val, Gln or Pro. In another aspect, the dispersinvariant comprises or consists of a substitution selected from the groupconsisting of: L170D, L170K, L170S, L170H, L170N, L170R, L170E, L170F,L170M, L170A, L170W, L170Y, L170V, L170Q and L170P, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 171 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 171 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Val, Ala, Trp, Arg, Cys, Glu, Lys, Tyr,Met, Gln or Leu. In another aspect, the dispersin variant comprises orconsists of a substitution selected from the group consisting of: D171V,D171A, D171W, D171R, D171C, D171E, D171K, D171Y, D171M, D171Q and D171L,wherein the position corresponds to the positions of SEQ ID NO: 1 andwherein the variant has at least at least 60%, at least 65%, at least70%, at least 75%, at least 80%, at least 85%, at least 90%, at least95%, such as at least 96%, at least 97%, at least 98%, or at least 99%,but less than 100%, sequence identity to the amino acid sequence shownin SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 173 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 173 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Cys. In another aspect, the dispersinvariant comprises or consists of the substitution I173C, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO: 1

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 174 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 174 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Trp, His, Met, Asn or Val. In anotheraspect, the dispersin variant comprises or consists of a substitutionselected from the group consisting of: D174W, D174H, D174M, D174N andD174V, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 175 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 175 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Tyr. In another aspect, the dispersinvariant comprises or consists of the substitution F175Y, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 177 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 177 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Met. In another aspect, the dispersinvariant comprises or consists of the substitution N177M, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of adeletion of the amino acid at a position corresponding to position 178of SEQ ID NO: 1. In another aspect, the dispersin variant comprises orconsists of a substitution at a position corresponding to position 178of SEQ ID NO: 1. In another aspect, the amino acid at a positioncorresponding to position 178 of SEQ ID NO: 1 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val, preferably with Arg, Trp, Ala or Lys. Inanother aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of: Q178R, Q178W, Q178Aand Q178K, wherein the position corresponds to the positions of SEQ IDNO: 1 and wherein the variant has at least at least 60%, at least 65%,at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 179 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 179 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Thr. In another aspect, the dispersinvariant comprises or consists of the substitution I179T, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 181 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 181 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Thr, Phe, Gln, Gly, Asn, Cys, Lys, Trp,Val, Asp, Thr or Pro. In another aspect, the dispersin variant comprisesor consists of a substitution selected from the group consisting of:S181T, S181F, S181Q, S181G, S181N, S181C, S181K, S181W, S181V, S181D,S181T and S181P, wherein the position corresponds to the positions ofSEQ ID NO: 1 and wherein the variant has at least at least 60%, at least65%, at least 70%, at least 75%, at least 80%, at least 85%, at least90%, at least 95%, such as at least 96%, at least 97%, at least 98%, orat least 99%, but less than 100%, sequence identity to the amino acidsequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 185 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 185 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Ala, Arg, Met, Val or Trp. In anotheraspect, the dispersin variant comprises or consists of a substitutionselected from the group consisting of: E185A, E185R, E185M, E185V andE185W, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 186 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 186 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asp, Glu, Asn, Gln, Leu, Trp, Val, Ile,Lys, Met, Arg or His. In another aspect, the dispersin variant comprisesor consists of a substitution selected from the group consisting of:S186D, S186E, S186N, S186Q, S186L, S186W, S186V, S186I, S186K, S186M,S186R and S186H, wherein the position corresponds to the positions ofSEQ ID NO: 1 and wherein the variant has at least at least 60%, at least65%, at least 70%, at least 75%, at least 80%, at least 85%, at least90%, at least 95%, such as at least 96%, at least 97%, at least 98%, orat least 99%, but less than 100%, sequence identity to the amino acidsequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 187 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 187 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Gly, Arg, Trp, Val, Cys, Asp or Ser. Inanother aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of: K187G, K187R, K187W,K187V, K187C, K187D and K187S, wherein the position corresponds to thepositions of SEQ ID NO: 1 and wherein the variant has at least at least60%, at least 65%, at least 70%, at least 75%, at least 80%, at least85%, at least 90%, at least 95%, such as at least 96%, at least 97%, atleast 98%, or at least 99%, but less than 100%, sequence identity to theamino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 188 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 188 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Pro. In another aspect, the dispersinvariant comprises or consists of the substitution Y188P, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 189 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 189 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with, Leu, Val or Trp. In another aspect, thedispersin variant comprises or consists of a substitution selected fromthe group consisting of: E189V, E189L and E189W, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 199 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 199 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Tyr, Met, Cys or Leu. In another aspect,the dispersin variant comprises or consists of a substitution selectedfrom the group consisting of: S199Y, S199M, S199C and S199L, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 200 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 200 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Trp, Asp, Lys, Arg, Met, Phe, Asn or Leu.In another aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of: E200W, E200D, E200K,E200R, E200M, E200F, E200N and E200L, wherein the position correspondsto the positions of SEQ ID NO: 1 and wherein the variant has at least atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100%, sequenceidentity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 203 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 203 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Arg, Thr, Pro, Cys, Leu, Ser, Met, Gln,Thr, Asp, Gly, Glu or Val. In another aspect, the dispersin variantcomprises or consists of a substitution selected from the groupconsisting of: A203R, A203W, A203P, A203C, A203L, A2035, A203M, A203Q,A203T, A203D, A203G, A203E and A203V, wherein the position correspondsto the positions of SEQ ID NO: 1 and wherein the variant has at least atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100%, sequenceidentity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 204 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 204 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Leu, Tyr, Val, Trp or Met. In anotheraspect, the dispersin variant comprises or consists of a substitutionselected from the group consisting of: N204L, N204Y, N204V, N204W andN204M, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 205 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 205 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Ile. In another aspect, the dispersinvariant comprises or consists of the substitution L205I, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 207 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 207 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Lys, Ser, Cys, Gly, Asn, Val, Arg, Gln,Glu, Trp or Ala. In another aspect, the dispersin variant comprises orconsists of a substitution selected from the group consisting of: D207S,D207C, D207G, D207N, D207V, D207R, D207Q, D207E, D207W, D207K and D207A,wherein the position corresponds to the positions of SEQ ID NO: 1 andwherein the variant has at least at least 60%, at least 65%, at least70%, at least 75%, at least 80%, at least 85%, at least 90%, at least95%, such as at least 96%, at least 97%, at least 98%, or at least 99%,but less than 100%, sequence identity to the amino acid sequence shownin SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 208 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 208 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Ala, Val, Cys, Trp, Asp, Thr, Gln, Gly orLeu. In another aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of: S208A, S208V, S208C,S208W, S208D, S208T, S208Q, S208G and S208L, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 210 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 210 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Thr. In another aspect, the dispersinvariant comprises or consists of the substitution S210T, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of adeletion of the amino acid at a position corresponding to position 215of SEQ ID NO: 1. In another aspect, the dispersin variant comprises orconsists of a substitution at a position corresponding to position 215of SEQ ID NO: 1. In another aspect, the amino acid at a positioncorresponding to position 215 of SEQ ID NO: 1 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val, preferably with Lys, Arg, Leu or Met. Inanother aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of: Q215K, Q215R, Q215Land Q215M, wherein the position corresponds to the positions of SEQ IDNO: 1 and wherein the variant has at least at least 60%, at least 65%,at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 217 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 217 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Val. In another aspect, the dispersinvariant comprises or consists of the substitution S217V, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 218 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 218 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Gln, Arg, Ala, Val or Leu. In anotheraspect, the dispersin variant comprises or consists of a substitutionselected from the group consisting of: T218Q, T218R, T218A, T218V andT218L, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 221 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 221 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asn. In another aspect, the dispersinvariant comprises or consists of the substitution S221N, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 222 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 222 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asp. In another aspect, the dispersinvariant comprises or consists of the substitution G222D, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 224 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 224 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Ala or Pro. In another aspect, thedispersin variant comprises or consists of the substitution E224A orE224P, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 225 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 225 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Gly. In another aspect, the dispersinvariant comprises or consists of the substitution S225G, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 227 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 227 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Ala, Gln, Arg, Ser, Thr or Lys. In anotheraspect, the dispersin variant comprises or consists of the substitutionN227A, N227Q, N227R, N227S, N227T or N227K, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of adeletion of an amino acid at a position corresponding to position 230 ofSEQ ID NO: 1. In another aspect, the dispersin variant comprises orconsists of a substitution at a position corresponding to position 230of SEQ ID NO: 1. In another aspect, the amino acid at a positioncorresponding to position 230 of SEQ ID NO: 1 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val, preferably with Asn, Arg, Trp or Thr. Inanother aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of: D230N, D230R, D230Wand D230T, wherein the position corresponds to the positions of SEQ IDNO: 1 and wherein the variant has at least at least 60%, at least 65%,at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 232 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 232 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asp or Val. In another aspect, thedispersin variant comprises or consists of the substitution E232D orE232V, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 233 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 233 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Glu, Asp, His, Gln, Arg or Trp. In anotheraspect, the dispersin variant comprises or consists of a substitutionselected from the group consisting of: N233E, N233D, N233H, N233Q, N233Rand N233W, wherein the position corresponds to the positions of SEQ IDNO: 1 and wherein the variant has at least at least 60%, at least 65%,at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 234 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 234 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Arg. In another aspect, the dispersinvariant comprises or consists of the substitution W234R, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 235 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 235 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Trp, Ala, Ser, Val, Leu, Met, Asn, His,Glu, Phe, Pro or Ile. In another aspect, the dispersin variant comprisesor consists of a substitution selected from the group consisting of:G235W, G235A, G235S, G235V, G235L, G235M, G235N, G235H, G235E, G235F,G235P and G235I, wherein the position corresponds to the positions ofSEQ ID NO: 1 and wherein the variant has at least at least 60%, at least65%, at least 70%, at least 75%, at least 80%, at least 85%, at least90%, at least 95%, such as at least 96%, at least 97%, at least 98%, orat least 99%, but less than 100%, sequence identity to the amino acidsequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 237 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 237 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Trp, Tyr, Gly, Met, Asn or Cys. In anotheraspect, the dispersin variant comprises or consists of a substitutionselected from the group consisting of: S237W, S237Y, S237G, S237M, S237Nand S237C, wherein the position corresponds to the positions of SEQ IDNO: 1 and wherein the variant has at least at least 60%, at least 65%,at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 244 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 244 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Glu, Met or Cys. In another aspect, thedispersin variant comprises or consists of a substitution selected fromthe group consisting of: Y244E, Y244M and Y244C, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 249 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 249 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Gln, Arg, Trp, His, Lys or Tyr. In anotheraspect, the dispersin variant comprises or consists of a substitutionselected from the group consisting of: L249Q, L249R, L249W, L249H, L249Kand L249Y, wherein the position corresponds to the positions of SEQ IDNO: 1 and wherein the variant has at least at least 60%, at least 65%,at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 251 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 251 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Ala, Leu, Arg, Trp or Asn. In anotheraspect, the dispersin variant comprises or consists of a substitutionselected from the group consisting of S251A, S251L, S251R, S251W andS251N, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 252 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 252 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Pro or Cys. In another aspect, thedispersin variant comprises or consists of the substitution N252P orN252C, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 253 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 253 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asp or Trp. In another aspect, thedispersin variant comprises or consists of the substitution G253D, orG253W, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 254 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 254 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Ile, Leu, Met or Tyr. In another aspect,the dispersin variant comprises or consists of a substitution selectedfrom the group consisting of F254I, F254L, F254M and F254Y, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 256 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 256 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Glu, Arg or Asp. In another aspect, thedispersin variant comprises or consists of the substitution Q256E, Q256Ror Q256D, wherein the position corresponds to the positions of SEQ IDNO: 1 and wherein the variant has at least at least 60%, at least 65%,at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of adeletion of the amino acid at a position corresponding to position 260of SEQ ID NO: 1. In another aspect, the dispersin variant comprises orconsists of a substitution at a position corresponding to position 260of SEQ ID NO: 1. In another aspect, the amino acid at a positioncorresponding to position 260 of SEQ ID NO: 1 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val, preferably with Gln, Leu, Cys, Arg, Met,Lys, Ala, Val, Ile, Thr, Glu, Trp or Tyr. In another aspect, thedispersin variant comprises or consists of a substitution selected fromthe group consisting of N260Q, N260L, N260C, N260R, N260M, N260K, N260A,N260V, N260I, N260T, N260E, N260W and N260Y, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 261 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 261 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Ala, Asp, Arg or Trp. In another aspect,the dispersin variant comprises or consists of a substitution selectedfrom the group consisting of E261A, E261D, E261R and E261W, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of adeletion of the amino acid at a position corresponding to position 262of SEQ ID NO: 1. In another aspect, the dispersin variant comprises orconsists of a substitution at a position corresponding to position 262of SEQ ID NO: 1. In another aspect, the amino acid at a positioncorresponding to position 262 of SEQ ID NO: 1 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val, preferably with His, Phe, Trp or Tyr. Inanother aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of Q262H, Q262F, Q262Wand Q262Y, wherein the position corresponds to the positions of SEQ IDNO: 1 and wherein the variant has at least at least 60%, at least 65%,at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 263 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 263 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Gln, Leu or Lys. In another aspect, thedispersin variant comprises or consists of a substitution selected fromthe group consisting of M263Q, M263L and M263K, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of adeletion of the amino acid at a position corresponding to position 264of SEQ ID NO: 1. In another aspect, the dispersin variant comprises orconsists of a substitution at a position corresponding to position 264of SEQ ID NO: 1. In another aspect, the amino acid at a positioncorresponding to position 264 of SEQ ID NO: 1 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val, preferably with Glu, Cys or Asn. In anotheraspect, the dispersin variant comprises or consists of a substitutionselected from the group consisting of D264E, D264C and D264N, whereinthe position corresponds to the positions of SEQ ID NO: 1 and whereinthe variant has at least at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 95%, suchas at least 96%, at least 97%, at least 98%, or at least 99%, but lessthan 100%, sequence identity to the amino acid sequence shown in SEQ IDNO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 265 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 265 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Phe. In another aspect, the dispersinvariant comprises or consists of the substitution Y265F, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 267 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 267 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Thr or Ser. In another aspect, thedispersin variant comprises or consists of the substitution N267T orN267S, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 268 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 268 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Glu, Arg, Cys or Met. In another aspect,the dispersin variant comprises or consists of a substitution selectedfrom the group consisting of W268E, W268R, W268C and W268M, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 270 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 270 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Phe. In another aspect, the dispersinvariant comprises or consists of the substitution Y270F, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 271 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 271 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Gly or Asp. In another aspect, thedispersin variant comprises or consists of the substitution A271G orA271D, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 272 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 272 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asp, Met, Pro, Ile, Val or Trp. In anotheraspect, the dispersin variant comprises or consists of a substitutionselected from the group consisting of H272D, H272M, H272P, H272I, H272Vand H272W wherein the position corresponds to the positions of SEQ IDNO: 1 and wherein the variant has at least at least 60%, at least 65%,at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 273 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 273 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Trp. In another aspect, the dispersinvariant comprises or consists of the substitution N273W, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 274 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 274 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with His, Arg or Ala. In another aspect, thedispersin variant comprises or consists of a substitution selected fromthe group consisting of K274H, K274R and K274A, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 276 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 276 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Ala, Asn, Lys, Cys, Gly, Leu, Met, Pro,Ser, Val or Trp. In another aspect, the dispersin variant comprises orconsists of a substitution selected from the group consisting of F276A,F276N, F276K, F276C, F276G, F276L, F276M, F276P, F276S, F276V and F276W,wherein the position corresponds to the positions of SEQ ID NO: 1 andwherein the variant has at least at least 60%, at least 65%, at least70%, at least 75%, at least 80%, at least 85%, at least 90%, at least95%, such as at least 96%, at least 97%, at least 98%, or at least 99%,but less than 100%, sequence identity to the amino acid sequence shownin SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 278 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 278 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Val, Gln, Ala, Asn or Lys. In anotheraspect, the dispersin variant comprises or consists of a substitutionselected from the group consisting of I278V, I278Q, I278A, 1278N andI278K, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 279 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 279 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Cys, Glu, Asn, Asp or Gly. In anotheraspect, the dispersin variant comprises or consists of a substitutionselected from the group consisting of S279C, S279E, S279N, S279D andS279G, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 280 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 280 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Glu or Cys. In another aspect, thedispersin variant comprises or consists of the substitution D280E orD280C, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of adeletion of the amino acid at a position corresponding to position 281of SEQ ID NO: 1. In another aspect, the dispersin variant comprises orconsists of a substitution at a position corresponding to position 281of SEQ ID NO: 1. In another aspect, the amino acid at a positioncorresponding to position 281 of SEQ ID NO: 1 is substituted with Ala,Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro,Ser, Thr, Trp, Tyr, or Val, preferably with His, Ala, Lys, Asn, Cys,Proor Arg. In another aspect, the dispersin variant comprises or consistsof a substitution selected from the group consisting of Y281H, Y281A,Y281K, Y281N, Y281C, Y281P and Y281R, wherein the position correspondsto the positions of SEQ ID NO: 1 and wherein the variant has at least atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100%, sequenceidentity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 282 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 282 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asn or Glu. In another aspect, thedispersin variant comprises or consists of a substitution selected fromthe group consisting of Y282N and Y282E, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 283 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 283 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Ile. In another aspect, the dispersinvariant comprises or consists of the substitution H283I, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 284 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 284 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Leu, Ile, Val, Asn, Pro or Thr. In anotheraspect, the dispersin variant comprises or consists of a substitutionselected from the group consisting of A284L, A284I, A284V, A284N, A284Pand A284T, wherein the position corresponds to the positions of SEQ IDNO: 1 and wherein the variant has at least at least 60%, at least 65%,at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 287 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 287 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asn. In another aspect, the dispersinvariant comprises or consists of the substitution T287N, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 288 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 288 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Pro, Asp, Asn or Lys. In another aspect,the dispersin variant comprises or consists of a substitution selectedfrom the group consisting of S288P, S288D, S288N and S288K, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 290 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 290 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Ile. In another aspect, the dispersinvariant comprises or consists of the substitution V290I, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 291 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 291 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Arg, Leu or Val. In another aspect, thedispersin variant comprises or consists of a substitution selected fromthe group consisting of K291R, K291L and K291V, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 296 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 296 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Cys. In another aspect, the dispersinvariant comprises or consists of the substitution T296C, wherein theposition corresponds to the positions of SEQ ID NO: 1 and wherein thevariant has at least at least 60%, at least 65%, at least 70%, at least75%, at least 80%, at least 85%, at least 90%, at least 95%, such as atleast 96%, at least 97%, at least 98%, or at least 99%, but less than100%, sequence identity to the amino acid sequence shown in SEQ ID NO:1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 300 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 300 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asp or Ala. In another aspect, thedispersin variant comprises or consists of the substitution E300D orE300A, wherein the position corresponds to the positions of SEQ ID NO: 1and wherein the variant has at least at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100%, sequence identity to the amino acid sequenceshown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 301 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 301 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asn, Cys or Arg. In another aspect, thedispersin variant comprises or consists of a substitution selected fromthe group consisting of H301N, H301C and H301R, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 303 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 303 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Gln, Ala, Cys, Lys, Gly, Trp or Arg. Inanother aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of T303Q, T303A, T303C,T303K, T303G, T303W and T303R, wherein the position corresponds to thepositions of SEQ ID NO: 1 and wherein the variant has at least at least60%, at least 65%, at least 70%, at least 75%, at least 80%, at least85%, at least 90%, at least 95%, such as at least 96%, at least 97%, atleast 98%, or at least 99%, but less than 100%, sequence identity to theamino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 304 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 304 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Met or Cys. In another aspect, thedispersin variant comprises or consists of a substitution selected fromthe group consisting of D304M and D304C, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 305 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 305 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Met or Asn. In another aspect, thedispersin variant comprises or consists of the L305M or L305N, whereinthe position corresponds to the positions of SEQ ID NO: 1 and whereinthe variant has at least at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 95%, suchas at least 96%, at least 97%, at least 98%, or at least 99%, but lessthan 100%, sequence identity to the amino acid sequence shown in SEQ IDNO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 306 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 306 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Cys. In another aspect, the dispersinvariant comprises or consists of the S306C, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 308 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 308 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Asp, Glu, Ala, Val, Gln, Ser, Tyr, Gly,Leu, Thr or Ile. In another aspect, the dispersin variant comprises orconsists of a substitution selected from the group consisting of K308D,K308E, K308A, K308V, K308Q, K308S, K308Y, K308G, K308L, K308T and K308I,wherein the position corresponds to the positions of SEQ ID NO: 1 andwherein the variant has at least at least 60%, at least 65%, at least70%, at least 75%, at least 80%, at least 85%, at least 90%, at least95%, such as at least 96%, at least 97%, at least 98%, or at least 99%,but less than 100%, sequence identity to the amino acid sequence shownin SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 309 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 309 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Glu, Gly, Cys, Leu, Asp, Gln, Asn, Thr,Ala, Ser, Met, His or Ile. In another aspect, the dispersin variantcomprises or consists of a substitution selected from the groupconsisting of K309E, K309G, K309C, K309L, K309D, K309Q, K309N, K309T,K309A, K309S, K309M, K309H and K309I, wherein the position correspondsto the positions of SEQ ID NO: 1 and wherein the variant has at least atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100%, sequenceidentity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 312 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 312 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Glu, Ala, Gln, Ser, Trp, Leu, Asn or Met.In another aspect, the dispersin variant comprises or consists of asubstitution selected from the group consisting of K312E, K312A, K312Q,K312S, K312W, K312L, K312N and K312M, wherein the position correspondsto the positions of SEQ ID NO: 1 and wherein the variant has at least atleast 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100%, sequenceidentity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 314 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 314 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Leu, Val or Ile. In another aspect, thedispersin variant comprises or consists of a substitution selected fromthe group consisting of E314L, E314V and E314I, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 315 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 315 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Ile or Val. In another aspect, thedispersin variant comprises or consists of the L315I or L315V, whereinthe position corresponds to the positions of SEQ ID NO: 1 and whereinthe variant has at least at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 95%, suchas at least 96%, at least 97%, at least 98%, or at least 99%, but lessthan 100%, sequence identity to the amino acid sequence shown in SEQ IDNO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 319 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 319 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Ala. In another aspect, the dispersinvariant comprises or consists of the R319A, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 321 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 321 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Phe. In another aspect, the dispersinvariant comprises or consists of the Y321F, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In another aspect, the dispersin variant comprises or consists of asubstitution at a position corresponding to position 323 of SEQ IDNO: 1. In another aspect, the amino acid at a position corresponding toposition 323 of SEQ ID NO: 1 is substituted with Ala, Arg, Asn, Asp,Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp,Tyr, or Val, preferably with Arg. In another aspect, the dispersinvariant comprises or consists of the N323R, wherein the positioncorresponds to the positions of SEQ ID NO: 1 and wherein the variant hasat least at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the amino acid sequence shown in SEQ ID NO: 1.

In one aspect, the variant comprises or consists of one or more thesubstitutions at a position selected from the group consisting of: 2, 3,12, 15, 17, 18, 19, 22, 23, 24, 25, 26, 30, 32, 34, 43, 44, 45, 49, 52,54, 56, 57, 58, 59, 60, 62, 63, 67, 68, 71, 72, 74, 77, 79, 80, 81, 82,90, 99, 100, 103, 104, 105, 106, 107, 110, 111, 113, 114, 116, 117, 118,119, 120, 122, 123, 124, 125, 126, 127, 128, 131, 135, 138, 139, 140,142, 145, 147, 148, 149, 150, 151, 152, 163, 164, 167, 168, 170, 171,173, 174, 175, 177, 178, 179, 181, 185, 186, 187, 188, 189, 199, 200,203, 204, 205, 207, 208, 210, 215, 217, 218, 221, 222, 224, 225, 227,230, 232, 233, 234, 235, 237, 244, 249, 251, 252, 253, 254, 256, 260,261, 262, 263, 264, 265, 267, 268, 270, 271, 272, 273, 274, 276, 278,279, 280, 281, 282, 283, 284, 287, 288, 290, 291, 296, 300, 301, 303,304, 305, 306, 308, 309, 312, 314, 315, 319, 321 and 323, wherein theposition corresponds to the positions of SEQ ID NO: 1, wherein thevariant has at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99% identity to the polypeptide shown in SEQ ID NO: 1 which hasdispersin, preferably beta-1,6 N-acetylglucosaminidase activity, andfurther the variant has at least one improved property compared to areference enzyme e.g. compared to SEQ ID NO: 1.

In one aspect, the variant comprises or consists of one or more thesubstitutions at a position selected from the group consisting of: 2, 3,12, 15, 17, 18, 22, 23, 24, 25, 30, 49, 56, 57, 59, 62, 63, 68, 72, 74,77, 82, 90, 99, 100, 106, 114, 123, 124, 125, 135, 138, 140, 163, 167,170, 171, 173, 174, 175, 178, 179, 181, 185, 186, 187, 188, 189, 199,203, 204, 205, 207, 210, 215, 218, 221, 225, 227, 227, 232, 235, 237,244, 252, 256, 260, 262, 263, 264, 265, 267, 270, 272, 273, 274, 276,278, 279, 280, 281, 282, 283, 284, 288, 290, 291, 296, 303, 304, 305,306, 308, 309, 312, 314, 315, 319, 321, 322 and 323, wherein theposition corresponds to the positions of SEQ ID NO: 1, wherein thevariant has at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99% identity to the polypeptide shown in SEQ ID NO: 1 which hasdispersin, preferably beta-1,6 N-acetylglucosaminidase activity, andfurther the variant has at least one improved property compared to areference enzyme e.g. compared to SEQ ID NO: 1.

In one aspect, the variant comprises or consists of one or more thesubstitutions at a position selected from the group consisting of: 31,33, 36, 73, 75, 94, 141, 144, 241, 277, 294, 297, 299, wherein theposition corresponds to the positions of SEQ ID NO: 1, wherein thevariant has at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99% identity to the polypeptide shown in SEQ ID NO: 1 which hasdispersin, preferably beta-1,6 N-acetylglucosaminidase activity, andfurther the variant has at least one improved property compared to areference enzyme e.g. compared to SEQ ID NO: 1.

In one aspect, the variant comprises or consists of one or more of thealterations selected from the group consisting of: D2A, D2L, D2N, D2R,D2V, D2W, Q3F, Q3I, Q3L, Q3M, Q3P, Q3V, Q3Y, Q3T, S12A, H15F, H15Y,T17W, T17C, T17E, T17F, T17M, T17R, T17V, V18L, E19D, E19K, E19N, E19P,K22A, K22M, K22V, S23A, S23C, S23E, S23I, S23L, S23R, S23T, S23V, L24I,V25R, D26M, Y30*, Y30A, Y30D, Y30L, Y30M, Y30N, Y30R, Y30T, Y30V, G32L,G32M, G32R, N34D, N43*, N43H, N43L, E44*, N45D, N45L, N45V, A49W, A49Y,Y52*, Y52M, G54L, G54M, G54N, S56T, S56W, S57W, E58N, N59A, N59C, N59D,N59E, N59F, N59M, N59R, N59V, N59W, T60V, N62C, N62D, N62H, N62Q, N62W,T63C, T63D, T63L, T63N, T63R, T63V, K67A, K67L, N68L, N68Q, L71H, L71N,L71R, L71V, L71W, S72*, S72C, S72D, S72E, S72F, S72G, S72I, S72M, S72N,S72R, S72T, S72Y, I74L, S77A, D79V, K80*, K80E, K80H, K80L, K80N, K80Q,K80R, K80V, K80W, D81A, D81G, D81L, D81N, D81R, D81S, D81T, D81V, D81W,I82V, L90F, E99Q, E99R, L100S, K103A, K103R, K103V, K104N, K104W, D105N,V106A, V106D, V106E, V106H, V106K, V106L, V106M, V106N, V106Q, V106R,V106W, V106Y, K107A, K107C, K107L, K107M, K107T, K107V, K107W, N110M,N110R, N110V, D111A, D111E, D111M, D111N, D111Q, D111R, D111V, D111W,V113T, T114C, T114S, Y116D, Y116N, Y116R, S117*, S117D, S117H, S117N,S117P, E118*, E118A, E118D, E118G, E118L, E119G, E119W, T120I, T120L,T120M, T120V, T120W, D122*, D122H, D122R, Y123W, Y124C, Y124H, Y124I,Y124K, Y124L, Y124M, Y124N, Y124Q, Y124R, Y124T, Y124V, Y124W, D125C,D125G, D125H, D125K, D125Q, D125R, N126V, R127D, R127H, R127K, R127L,R127M, R127Q, R127W, V128A, V128C, V128D, V128L, V128T, D131V, Q135*,Q135A, Q135D, Q135E, Q135K, Q135M, Q135Y, D138K, D138L, D138M, D138N,D138Q, D138R, D138S, D138V, D138W, E139W, V140I, D142R, D142W, Y145*,Y145H, Y145L, Y145N, Y145V, P147A, P147C, P147D, P147F, P147G, P147L,P147M, P147R, P147S, P147T, P147V, K148A, K148D, K148L, K148V, F149L,F149M, F149N, E150A, E150D, E150H, E150K, E150L, E150M, E150N, E150R,E150V, E150W, E150Y, G151A, G151C, G151D, G151L, G151N, G151P, G151S,G151W, K152D, K152L, K152R, S163P, G164D, G164E, G164H, G164S, G164V,V167A, V167D, V167E, V167L, V167P, V167Q, V167R, V167W, H168N, L170A,L170D, L170E, L170F, L170H, L170K, L170M, L170N, L170P, L170Q, L170R,L170S, L170V, L170W, L170Y, D171A, D171C, D171E, D171K, D171L, D171M,D171Q, D171R, D171V, D171W, D171Y, I173C, D174H, D174M, D174N, D174V,D174W, F175Y, N177M, Q178*, Q178A, Q178K, Q178R, Q178W, I179T, S181C,S181D, S181F, S181G, S181K, S181N, S181P, S181Q, S181T, S181V, S181W,E185A, E185M, E185R, E185V, E185W, S186D, S186E, S186H, S186I, S186K,S186L, S186M, S186N, S186Q, S186R, S186V, S186W, K187C, K187D, K187G,K187R, K187S, K187V, K187W, Y188P, E189L, E189V, E189W, S199C, S199L,S199M, S199Y, E200D, E200F, E200K, E200L, E200M, E200N, E200R, E200W,A203C, A203D, A203E, A203G, A203L, A203M, A203P, A203Q, A203R, A203S,A203T, A203V, A203W, N204L, N204M, N204V, N204W, N204Y, L205I, D207A,D207C, D207E, D207G, D207K, D207N, D207Q, D207R, D207S, D207V, D207W,S208A, S208C, S208D, S208G, S208L, S208Q, S208T, S208V, S208W, S210T,Q215K, Q215R, Q215M, Q215L, Q215*, S217V, T218A, T218L, T218Q, T218R,T218V, S221N, G222D, E224A, E224P, S225G, N227A, N227Q, N227R, N227S,N227T, N227K, D230*, D230N, D230R, D230T, D230W, E232D, E232V, N233D,N233E, N233H, N233Q, N233R, N233W, W234R, G235W, G235A, G235E, G235F,G235H, G235I, G235L, G235M, G235N, G235P, G235S, G235V, S237C, S237G,S237M, S237N, S237W, S237Y, Y244C, Y244E, Y244M, L249H, L249K, L249Q,L249R, L249W, L249Y, S251A, S251L, S251N, S251R, S251W, N252P, N252C,G253D, G253W, F254I, F254L, F254M, F254Y, Q256D, Q256E, Q256R, N260*,N260A, N260C, N260E, N260I, N260K, N260L, N260M, N260Q, N260R, N260T,N260V, N260W, N260Y, E261*, E261A, E261D, E261R, E261W, Q262*, Q262F,Q262H, Q262W, Q262Y, M263K, M263L, M263Q, D264*, D264C, D264E, D264N,Y265F, N267S, N267T, W268C, W268E, W268M, W268R, Y270F, A271D, A271G,H272D, H272I, H272M, H272P, H272V, H272W, N273W, K274R, K274A, K274H,F276A, F276C, F276K, F276N, F276G, F276L, F276M, F276P, F276S, F276V,F276W, I278A, I278K, I278N, I278Q, I278V, S279C, S279D, S279E, S279G,S279N, D280C, D280E, Y281*, Y281A, Y281C, Y281H, Y281K, Y281N, Y281P,Y281R, Y282E, Y282N, H283I, A284I, A284L, A284N, A284P, A284T, A284V,T287N, S288P, S288D, S288K, S288N, V290I, K291L, K291R, K291V, T296C,E300A, E300D, H301C, H301N, H301R, T303A, T303C, T303G, T303K, T303Q,T303R, T303W, D304C, D304M, L305M, L305N, S306C, K308A, K308D, K308E,K308G, K308I, K308L, K308Q, K308S, K308T, K308V, K308Y, K309A, K309C,K309D, K309E, K309G, K309H, K309L, K309M, K309N, K309Q, K309S, K309T,K309I, K312A, K312E, K312L, K312M, K312N, K312Q, K312S, K312W, E314I,E314L, E314V, L315I, L315V, R319A, Y321F and N323R, wherein the positioncorresponds to the positions of SEQ ID NO: 1, wherein the variant has atleast 65%, at least 70%, at least 75%, at least 80%, at least 85%, atleast 90%, at least 91%, at least 92%, at least 93%, at least 94%, atleast 95%, at least 96%, at least 97%, at least 98%, at least 99%identity to the polypeptide shown in SEQ ID NO: 1 which has dispersin,prefereably beta-1,6 N-acetylglucosaminidase activity, and further thevariant has at least one improved property compared to a referenceenzyme e.g. compared to SEQ ID NO: 1.

In one aspect, the variant comprises or consists of one or more of thesubstitutions selected from the group consisting of: D2V, Q3F, Q3I,S12A, H15Y, T17W, T17E, V18L, K22M, S23I, S23C, S23T, S23L, S23V, S23E,V25R, Y30L, A49W, A49Y, S56T, N59D, N59C, N59E, N59R, N59F, N59W, N59V,N62C, N62D, T63C, N68Q, S72D, S72E, I74L, S77A, I82V, L90F, E99Q, L100S,T114S, T114C, Y123W, Y124I, Y124M, Y124R, Y124V, Y124Q, Y124T, Y124K,D125R, D125C, D125G, D125K, D125Q, Q135M, D138R, D138K, D138Q, L170D,L170K, L170S, L170H, D171E, D171K, D171Y, D171M, D171Q, D171L, I173C,D174W, D174H, D174M, D174N, F175Y, Q178K, I179T, S181T, S181F, S181Q,S181G, S181N, S181C, E185R, E185M, E185V, S186K, S186M, S186R, S186H,K187G, Y188P, E189V, S199C, S199L, A203G, A203E, A203V, N204L, N204Y,N204V, L205I, D207N, D207S, D207C, D207G, S210T, Q215R, T218Q, N227T,N227K, E232D, G235W, S237W, Y244M, Y244C, N252P, N252C, Q256E, Q262H,N260Q, M263Q, D264E, Y265F, N267T, N267S, Y270F, H272M, H272P, H272I,H272V, N273W, K274H, F276A, F276N, F276K, F276C, I278V, S279N, S279D,S279G, D280E, D280C, Y281P, Y282N, H283I, A284T, A284L, A284I, S288K,V290I, K291R, T296C, T303Q, D304M, D304C, L305M, L305N, S306C, K308D,K308A, K308V, K308Q, K308S, K308Y, K308G, K308L, K308T, K308I, K309C,K309L, K309D, K309Q, K309N, K309T, K309A, K309S, K309M, K309H, K312A,K312Q, K312S, K312W, K312L, K312N, E314L, E314V, E314I, L315I, R319A andN323R, wherein the position corresponds to the positions of SEQ ID NO:1, wherein the variant has at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99% identity to the polypeptide shown in SEQ ID NO:1 which has dispersin, preferably beta-1,6 N-acetylglucosaminidaseactivity, and further the variant has at least one improved propertycompared to a reference enzyme e.g. compared to SEQ ID NO: 1 or comparedto SEQ ID NO: 15.

In some aspects, the improved property is increased stability e.g.improved detergent stability, improved in-wash stability and improvedthermostability. Some aspects of the invention relate to dispersinvariants having an improvement factor above 1 (e.g. measured as HIF),when the dispersin variants are tested for a property of interest in arelevant assay, wherein the property of the reference dispersin is givena value of 1. In some aspects, the property is stability, such asstorage stability. Some aspects of the invention relate to dispersinvariants having an improvement factor above 1 (e.g. measured as HIF),when the dispersin variants are tested for a property of interest in theassay described in Example 2, 3, 4, 5 or 6, wherein the property of thereference dispersin is given a value of 1. In some aspects, the propertyis stability, such as storage stability.

In some aspects, the improved property is increased stability e.g.improved detergent stability, improved in-wash stability and improvedthermostability. Some aspects of the invention relate to dispersinvariants having an improvement factor above 1 when the dispersinvariants are tested for a property of interest in a relevant assay,wherein the property of the reference dispersin is given a value of 1.In some aspects, the property is stability, such as storage stability.

In some aspects, the improved property is improved detergent stability.

In some aspects, the improved property is improved protease stability.

In some aspects a variant according to the invention is improved underthe measured conditions when the residual activity ratio, defined as

${{Residual}{Activity}{{Ratio}{}({RAR})}} = \frac{{RA}_{variant}}{{RA}_{reference}}$

is above 1 compared to the reference dispersin, as shown in Examples 2aand 3. In some aspects, the variant is improved compared to thereference dispersin.

In some aspects, the variants according to the invention have improvedstability relative to a reference dispersin measured as a residualactivity ratio (RAR) that is greater than 1.0.

The Residual Activity Ratio (RAR) for the dispersin variants of theinvention may be calculated as RA_(variant)/RA_(reference). Improvedvariants were identified as variants having residual activity ratio(RAR) larger than 1.0 compared to the reference dispersin, as shown inExample 2a and 3.

In some aspects, the variants according to the invention have improvedstability relative to a reference dispersin measured as an Half-lifeImprovement Factor (HIF) that is greater than 1.0.

The Half-life Improvement Factor (HIF) for the dispersin variants of theinvention may be calculated as T_(1/2variant)/T_(1/2reference). Improvedvariants were identified as variants having a Half-life ImprovementFactor HIF larger than 1.0 compared to the reference dispersin, as shownin Example 2b. Improved alterations e.g. substitutions according to theinvention are those which result in an improvement factor e.g. HIF orRAR above 1 or is a variant which have at least one improved propertycompared to the starting molecule i.e. the precursor, reference parentpolypeptide etc. In a particular preferred aspect, dispersin variantscomprising the alterations according to the invention results indispersin variants having improved stability where HIF>1.0. In someaspects, the variants according to the invention have a Half-lifeImprovement Factor (HIF) which is at least 1.1; 1.2; 1.3; 1.4; 1.5; 1.6;1.7; 1.8; 1.9; 2.0; 2.1; 2.2; 2.3; 2.4; 2.5; 2.6; 2.7, 2.8; 2.9; 3.0,3.1; 3.2; 3.3; 3.4; 3.5, 3.6, 3.7, 3.8, 3.9; 4.0, 4.1; 4.2; 4.3; 4.4;4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1; 5.2; 5.3; 5.4; 5.5, 5.6, 5.7, 5.8,5.9; 3.0, 6.1; 6.2; 6.3; 6.4; 6.5, 6.6, 6.7, 6.8, 6.9; 7.0, 7.1; 7.2;7.3; 7.4; 7.5, 7.6, 7.7, 7.8, 7.9; 8.0, 8.1; 8.2; 8.3; 8.4; 8.5, 8.6,8.7, 8.8, 8.9; 9.0, 9.1; 9.2; 9.3; 9.4; 9.5, 9.6, 9.7, 9.8, 9.9; 10.0,10.1; 10.2; 10.3; 10.4; 10.5, 10.6, 10.7, 10.8, 10.9; 12, 15, 16, 20, 25or 30 compared to a reference dispersin e.g. SEQ ID NO: 1 or compared toSEQ ID NO: 15.

In a preferred aspect, the combination of mutations according to theinvention results in dispersin variants having improved stability, whereRAR>1.0. In some aspects, the variants according to the invention have aResidual Activity Ratio (RAR) which is at least 1.1; 1.2; 1.3; 1.4; 1.5;1.6; 1.7; 1.8; 1.9; 2.0; 2.1; 2.2; 2.3; 2.4; 2.5; 2.6; 2.7, 2.8; 2.9;3.0, 3.1; 3.2; 3.3; 3.4; 3.5, 3.6, 3.7, 3.8, 3.9; 4.0, 4.1; 4.2; 4.3;4.4; 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1; 5.2; 5.3; 5.4; 5.5, 5.6, 5.7,5.8, 5.9; 3.0, 6.1; 6.2; 6.3; 6.4; 6.5, 6.6, 6.7, 6.8, 6.9; 7.0, 7.1;7.2; 7.3; 7.4; 7.5, 7.6, 7.7, 7.8, 7.9; 8.0, 8.1; 8.2; 8.3; 8.4; 8.5,8.6, 8.7, 8.8, 8.9; 9.0, 9.1; 9.2; 9.3; 9.4; 9.5, 9.6, 9.7, 9.8, 9.9;10.0, 10.1; 10.2; 10.3; 10.4; 10.5, 10.6, 10.7, 10.8, 10.9; 12, 15, 16,20, 25 or 30 compared to a reference dispersin e.g. SEQ ID NO: 1 orcompared to SEQ ID NO: 15.

One preferred embodiment relates to a dispersin variant having improvedstability, wherein HIF>1.0, compared to SEQ ID NO: 1 or compared to SEQID NO: 15. One preferred embodiment relates to a dispersin varianthaving improved stability, wherein the Half-life Improvement Factor isat least 1.5, compared to SEQ ID NO: 1 or compared to SEQ ID NO: 15,when measured as described in Example 2a.

One preferred embodiment relates to a dispersin variant having improvedstability, wherein RAR>1.0, compared to SEQ ID NO: 1 or compared to SEQID NO: 15. One preferred embodiment relates to a dispersin varianthaving improved stability, wherein the residual activity ratio (RAR) isat least 1.5, compared to SEQ ID NO: 1 or compared to SEQ ID NO: 15,when measured as described in Example 2a and 3.

One preferred embodiment relates to a dispersin variant having improvedstability, wherein HIF>1.0 compared to SEQ ID NO: 1. One preferredembodiment relates to a dispersin variant having improved stability,wherein the Half-life Improvement Factor (HIF) is at least 1.2 e.g. 1.5,compared to SEQ ID NO: 1 or compared to SEQ ID NO: 15, wherein HIF iscalculated as T_(1/2dispersinvariant)/T_(1/2reference) and the half-life(T_(1/) in minutes) for the dispersin variants and the dispersinreference (e.g. SEQ ID NO: 1 or 15) is calculated as: 20minutes×LN(0.5)/LN(RA) and the residual activity (RA) for each dispersinvariant and the reference dispersin (e.g. SEQ ID NO: 1 or 15) iscalculated as: slope (stress sample, 56° C. for 60 min)/slope(unstressedsample, 21° C. for 60 min), e.g. as described in Example 2b.

One preferred embodiment relates to a dispersin variant having improvedstability, wherein residual activity ratio (RAR)>1.0 compared to SEQ IDNO: 1 or compared to SEQ ID NO: 15. One preferred embodiment relates toa dispersin variant having improved stability, wherein the residualactivity ratio (RAR) is at least 1.2 e.g. 1.5, compared to SEQ ID NO: 1or compared to SEQ ID NO: 15, wherein RAR is calculated as,

${{Residual}{Activity}{Ratio}({RAR})} = \frac{{RA}_{variant}}{{RA}_{reference}}$

as described in 2a and 3, in the Example section.

One preferred aspect of the invention relates to a dispersin variant,which compared to a dispersin with SEQ ID NO: 1, comprises an alteratione.g. substitution at a position selected from the group consisting of:2, 3, 12, 15, 17, 18, 19, 22, 23, 24, 25, 26, 30, 32, 34, 43, 44, 45,49, 52, 54, 56, 57, 58, 59, 60, 62, 63, 67, 68, 71, 72, 74, 77, 79, 80,81, 82, 90, 99, 100, 103, 104, 105, 106, 107, 110, 111, 113, 114, 116,117, 118, 119, 120, 122, 123, 124, 125, 126, 127, 128, 131, 135, 138,139, 140, 142, 145, 147, 148, 149, 150, 151, 152, 163, 164, 167, 168,170, 171, 173, 174, 175, 177, 178, 179, 181, 185, 186, 187, 188, 189,199, 200, 203, 204, 205, 207, 208, 210, 215, 217, 218, 221, 222, 224,225, 227, 230, 232, 233, 234, 235, 237, 244, 249, 251, 252, 253, 254,256, 260, 261, 262, 263, 264, 265, 267, 268, 270, 271, 272, 273, 274,276, 278, 279, 280, 281, 282, 283, 284, 287, 288, 290, 291, 296, 300,301, 303, 304, 305, 306, 308, 309, 312, 314, 315, 319, 321 and 323,wherein the positions correspond to the positions of SEQ ID NO: 1(numbering according to SEQ ID NO: 1), wherein the variant has asequence identity to the polypeptide shown in SEQ ID NO: 1 or thepolypeptide sequence shown in SEQ ID NO: 15 of at least 65%, at least70%, at least 75%, at least 80%, at least 85%, at least 90%, at least91%, at least 92%, at least 93%, at least 94%, at least 95%, at least96%, at least 97%, at least 98% or at least 99% but less than 100%,wherein the variant has hexosaminidase e.g. beta-1,6N-acetylglucosaminidase activity and wherein the variant has improvedstability, measured as having an improvement factor e.g. RAR or HIF>1.0,compared to the polypeptide sequence shown in SEQ ID NO: 1 or comparedto the polypeptide sequence shown in SEQ ID NO: 15.

One preferred aspect of the invention relates to a dispersin variant,which compared to a dispersin with SEQ ID NO: 1, comprises an alteratione.g. substitution at a position selected from the group consisting of:2, 3, 12, 15, 17, 18, 22, 23, 24, 25, 30, 49, 56, 57, 59, 62, 63, 68,72, 74, 77, 82, 90, 99, 100, 106, 114, 123, 124, 125, 135, 138, 140,163, 167, 170, 171, 173, 174, 175, 178, 179, 181, 185, 186, 187, 188,189, 199, 203, 204, 205, 207, 210, 215, 218, 221, 225, 227, 227, 232,235, 237, 244, 252, 256, 260, 262, 263, 264, 265, 267, 270, 272, 273,274, 276, 278, 279, 280, 281, 282, 283, 284, 288, 290, 291, 296, 303,304, 305, 306, 308, 309, 312, 314, 315, 319, 321, 322 and 323, whereinthe positions correspond to the positions of SEQ ID NO: 1 (numberingaccording to SEQ ID NO: 1), wherein the variant has a sequence identityto the polypeptide shown in SEQ ID NO: 1 or the polypeptide sequenceshown in SEQ ID NO: 15 of at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98% or at least 99% but less than 100%, wherein the variant hashexosaminidase e.g. beta-1,6 N-acetylglucosaminidase activity andwherein the variant has improved stability, measured as having animprovement factor e.g. RAR or HIF>1.0, compared to the polypeptidesequence shown in SEQ ID NO: 1 or compared to the polypeptide sequenceshown in SEQ ID NO: 15.

One preferred aspect of the invention relates to a dispersin variant,which compared to a dispersin with SEQ ID NO: 1, comprises an alteratione.g. substitution at a position selected from the group consisting of:31, 33, 36, 73, 75, 94, 141, 144, 241, 277, 294, 297, 299, wherein thepositions correspond to the positions of SEQ ID NO: 1 (numberingaccording to SEQ ID NO: 1), wherein the variant has a sequence identityto the polypeptide shown in SEQ ID NO: 1 or the polypeptide sequenceshown in SEQ ID NO: 15 of at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98% or at least 99% but less than 100%, wherein the variant hashexosaminidase e.g. beta-1,6 N-acetylglucosaminidase activity andwherein the variant has improved stability, measured as having animprovement factor e.g. RAR or HIF>1.0, compared to the polypeptidesequence shown in SEQ ID NO: 1 or compared to the polypeptide sequenceshown in SEQ ID NO: 15.

One preferred aspect of the invention relates to a dispersin variant,which compared to a dispersin with SEQ ID NO: 1, comprises one or morealteration e.g. substitution selected from the group consisting of: D2A,D2L, D2N, D2R, D2V, D2W, Q3F, Q3I, Q3L, Q3M, Q3P, Q3V, Q3Y, Q3T, S12A,H15F, H15Y, T17W, T17C, T17E, T17F, T17M, T17R, T17V, V18L, E19D, E19K,E19N, E19P, K22A, K22M, K22V, S23A, S23C, S23E, S23I, S23L, S23R, S23T,S23V, L24I, V25R, D26M, Y30*, Y30A, Y30D, Y30L, Y30M, Y30N, Y30R, Y30T,Y30V, G32L, G32M, G32R, N34D, N43*, N43H, N43L, E44*, N45D, N45L, N45V,A49W, A49Y, Y52*, Y52M, G54L, G54M, G54N, S56T, S56W, S57W, E58N, N59A,N59C, N59D, N59E, N59F, N59M, N59R, N59V, N59W, T60V, N62C, N62D, N62H,N62Q, N62W, T63C, T63D, T63L, T63N, T63R, T63V, K67A, K67L, N68L, N68Q,L71H, L71N, L71R, L71V, L71W, S72*, S72C, S72D, S72E, S72F, S72G, S72I,S72M, S72N, S72R, S72T, S72Y, I74L, S77A, D79V, K80*, K80E, K80H, K80L,K80N, K80Q, K80R, K80V, K80W, D81A, D81G, D81L, D81N, D81R, D81S, D81T,D81V, D81W, I82V, L90F, E99Q, E99R, L100S, K103A, K103R, K103V, K104N,K104W, D105N, V106A, V106D, V106E, V106H, V106K, V106L, V106M, V106N,V106Q, V106R, V106W, V106Y, K107A, K107C, K107L, K107M, K107T, K107V,K107W, N110M, N110R, N110V, D111A, D111E, D111M, D111N, D111Q, D111R,D111V, D111W, V113T, T114C, T114S, Y116D, Y116N, Y116R, S117*, S117D,S117H, S117N, S117P, E118*, E118A, E118D, E118G, E118L, E119G, E119W,T120I, T120L, T120M, T120V, T120W, D122*, D122H, D122R, Y123W, Y124C,Y124H, Y124I, Y124K, Y124L, Y124M, Y124N, Y124Q, Y124R, Y124T, Y124V,Y124W, D125C, D125G, D125H, D125K, D125Q, D125R, N126V, R127D, R127H,R127K, R127L, R127M, R127Q, R127W, V128A, V128C, V128D, V128L, V128T,D131V, Q135*, Q135A, Q135D, Q135E, Q135K, Q135M, Q135Y, D138K, D138L,D138M, D138N, D138Q, D138R, D138S, D138V, D138W, E139W, V140I, D142R,D142W, Y145*, Y145H, Y145L, Y145N, Y145V, P147A, P147C, P147D, P147F,P147G, P147L, P147M, P147R, P147S, P147T, P147V, K148A, K148D, K148L,K148V, F149L, F149M, F149N, E150A, E150D, E150H, E150K, E150L, E150M,E150N, E150R, E150V, E150W, E150Y, G151A, G151C, G151D, G151L, G151N,G151P, G151S, G151W, K152D, K152L, K152R, S163P, G164D, G164E, G164H,G164S, G164V, V167A, V167D, V167E, V167L, V167P, V167Q, V167R, V167W,H168N, L170A, L170D, L170E, L170F, L170H, L170K, L170M, L170N, L170P,L170Q, L170R, L170S, L170V, L170W, L170Y, D171A, D171C, D171E, D171K,D171L, D171M, D171Q, D171R, D171V, D171W, D171Y, I173C, D174H, D174M,D174N, D174V, D174W, F175Y, N177M, Q178*, Q178A, Q178K, Q178R, Q178W,I179T, S181C, S181D, S181F, S181G, S181K, S181N, S181P, S181Q, S181T,S181V, S181W, E185A, E185M, E185R, E185V, E185W, S186D, S186E, S186H,S186I, S186K, S186L, S186M, S186N, S186Q, S186R, S186V, S186W, K187C,K187D, K187G, K187R, K187S, K187V, K187W, Y188P, E189L, E189V, E189W,S199C, S199L, S199M, S199Y, E200D, E200F, E200K, E200L, E200M, E200N,E200R, E200W, A203C, A203D, A203E, A203G, A203L, A203M, A203P, A203Q,A203R, A203S, A203T, A203V, A203W, N204L, N204M, N204V, N204W, N204Y,L205I, D207A, D207C, D207E, D207G, D207K, D207N, D207Q, D207R, D207S,D207V, D207W, S208A, S208C, S208D, S208G, S208L, S208Q, S208T, S208V,S208W, S210T, Q215K, Q215R, Q215M, Q215L, Q215*, S217V, T218A, T218L,T218Q, T218R, T218V, S221N, G222D, E224A, E224P, S225G, N227A, N227Q,N227R, N227S, N227T, N227K, D230*, D230N, D230R, D230T, D230W, E232D,E232V, N233D, N233E, N233H, N233Q, N233R, N233W, W234R, G235W, G235A,G235E, G235F, G235H, G235I, G235L, G235M, G235N, G235P, G235S, G235V,S237C, S237G, S237M, S237N, S237W, S237Y, Y244C, Y244E, Y244M, L249H,L249K, L249Q, L249R, L249W, L249Y, S251A, S251L, S251N, S251R, S251W,N252P, N252C, G253D, G253W, F254I, F254L, F254M, F254Y, Q256D, Q256E,Q256R, N260*, N260A, N260C, N260E, N260I, N260K, N260L, N260M, N260Q,N260R, N260T, N260V, N260W, N260Y, E261*, E261A, E261D, E261R, E261W,Q262*, Q262F, Q262H, Q262W, Q262Y, M263K, M263L, M263Q, D264*, D264C,D264E, D264N, Y265F, N267S, N267T, W268C, W268E, W268M, W268R, Y270F,A271D, A271G, H272D, H272I, H272M, H272P, H272V, H272W, N273W, K274R,K274A, K274H, F276A, F276C, F276K, F276N, F276G, F276L, F276M, F276P,F276S, F276V, F276W, I278A, I278K, I278N, I278Q, I278V, S279C, S279D,S279E, S279G, S279N, D280C, D280E, Y281*, Y281A, Y281C, Y281H, Y281K,Y281N, Y281P, Y281R, Y282E, Y282N, H283I, A284I, A284L, A284N, A284P,A284T, A284V, T287N, S288P, S288D, S288K, S288N, V290I, K291L, K291R,K291V, T296C, E300A, E300D, H301C, H301N, H301R, T303A, T303C, T303G,T303K, T303Q, T303R, T303W, D304C, D304M, L305M, L305N, S306C, K308A,K308D, K308E, K308G, K308I, K308L, K308Q, K308S, K308T, K308V, K308Y,K309A, K309C, K309D, K309E, K309G, K309H, K309L, K309M, K309N, K309Q,K309S, K309T, K309I, K312A, K312E, K312L, K312M, K312N, K312Q, K312S,K312W, E314I, E314L, E314V, L315I, L315V, R319A, Y321F and N323R,wherein the positions correspond to the positions of SEQ ID NO: 1(numbering according to SEQ ID NO: 1), wherein the variant has asequence identity to the polypeptide shown in SEQ ID NO: 1 or to thepolypeptide sequence shown in SEQ ID NO: 15 of at least 65%, at least70%, at least 75%, at least 80%, at least 85%, at least 90%, at least91%, at least 92%, at least 93%, at least 94%, at least 95%, at least96%, at least 97%, at least 98% or at least 99% but less than 100%,wherein the variant has hexosaminidase e.g. beta-1,6N-acetylglucosaminidase activity and wherein the variant has improvedstability, measured as having an improvement factor e.g. RAR or HIF>1.0,compared to the polypeptide sequence shown in SEQ ID NO: 1 or comparedto the polypeptide sequence shown in SEQ ID NO: 15.

One preferred aspect of the invention relates to a dispersin variant,which compared to a dispersin with SEQ ID NO: 1, comprises one or morealteration e.g. substitution selected from the group consisting of: D2A,D2L, D2N, D2R, D2V, D2W, Q3F, Q3I, Q3L, Q3M, Q3P, Q3V, Q3Y, Q3T, S12A,H15F, H15Y, T17W, T17C, T17E, T17F, T17M, T17R, T17V, V18L, E19D, E19N,E19P, K22A, K22M, K22V, S23C, S23E, S23I, S23L, S23R, S23T, S23V, V25R,D26M, Y30*, Y30D, Y30L, Y30M, Y30N, Y30R, Y30T, Y30V, G32L, G32M, G32R,N43*, N43H, N43L, E44*, N45D, N45L, N45V, A49W, A49Y, Y52*, Y52M, G54L,G54M, G54N, S56T, S56W, S57W, E58N, N59A, N59C, N59D, N59E, N59F, N59M,N59R, N59V, N59W, T60V, N62C, N62D, N62H, N62Q, N62W, T63C, T63D, T63L,T63N, T63R, T63V, K67A, K67L, N68L, N68Q, L71H, L71N, L71R, L71V, L71W,S72*, S72C, S72D, S72E, S72F, S72G, S72I, S72M, S72N, S72R, S72T, S72Y,I74L, S77A, D79V, K80*, K80E, K80H, K80L, K80N, K80Q, K80V, K80W, D81A,D81G, D81L, D81R, D81S, D81T, D81V, D81W, I82V, L90F, E99Q, E99R, L100S,K103A, K103R, K104N, K104W, D105N, V106A, V106D, V106E, V106H, V106K,V106L, V106M, V106N, V106Q, V106W, V106Y, K107A, K107C, K107L, K107M,K107T, K107V, K107W, N110M, N110R, N110V, D111A, D111E, D111M, D111N,D111Q, D111R, D111V, D111W, V113T, T114C, T114S, Y116D, Y116N, Y116R,S117*, S117D, S117H, S117N, S117P, E118*, E118A, E118D, E118G, E118L,E119G, E119W, T120I, T120L, T120M, T120V, T120W, D122*, D122H, D122R,Y123W, Y124C, Y124I, Y124K, Y124L, Y124M, Y124Q, Y124R, Y124T, Y124V,Y124W, D125C, D125G, D125K, D125Q, D125R, N126V, R127D, R127H, R127K,R127L, R127M, R127Q, R127W, V128C, V128L, V128T, D131V, Q135*, Q135A,Q135D, Q135E, Q135K, Q135M, Q135Y, D138K, D138L, D138M, D138Q, D138R,D138S, D138V, D138W, E139W, D142R, D142W, Y145*, Y145H, Y145L, Y145N,Y145V, P147A, P147C, P147D, P147F, P147G, P147L, P147M, P147R, P147S,P147T, P147V, K148A, K148D, K148L, K148V, F149L, F149M, F149N, E150D,E150H, E150K, E150L, E150M, E150N, E150R, E150V, E150W, E150Y, G151A,G151C, G151D, G151L, G151N, G151P, G151S, G151W, K152D, K152L, K152R,G164D, G164E, G164H, G164S, G164V, V167D, V167E, V167L, V167P, V167Q,V167R, V167W, H168N, L170A, L170D, L170E, L170F, L170H, L170K, L170M,L170N, L170P, L170Q, L170R, L170S, L170V, L170W, L170Y, D171A, D171C,D171E, D171K, D171L, D171M, D171Q, D171R, D171V, D171W, D171Y, I173C,D174H, D174M, D174N, D174V, D174W, F175Y, N177M, Q178*, Q178A, Q178K,Q178R, Q178W, I179T, S181C, S181D, S181F, S181G, S181N, S181P, S181Q,S181T, S181V, S181W, E185M, E185R, E185V, E185W, S186D, S186E, S186H,S186I, S186K, S186L, S186M, S186N, S186Q, S186R, S186V, S186W, K187C,K187D, K187G, K187R, K187S, K187V, K187W, Y188P, E189L, E189V, E189W,S199C, S199L, S199M, S199Y, E200D, E200F, E200K, E200L, E200M, E200N,E200R, E200W, A203C, A203D, A203E, A203G, A203L, A203M, A203P, A203R,A203S, A203T, A203V, A203W, N204L, N204M, N204V, N204W, N204Y, L205I,D207A, D207C, D207E, D207G, D207N, D207Q, D207S, D207V, D207W, S208A,S208C, S208D, S208G, S208L, S208Q, S208T, S208V, S208W, S210T, Q215R,Q215M, Q215L, Q215*, S217V, T218A, T218L, T218Q, T218R, T218V, G222D,E224A, E224P, N227A, N227Q, N227R, N227S, N227T, N227K, D230*, D230R,D230T, D230W, E232D, E232V, N233H, N233Q, N233R, N233W, W234R, G235W,G235A, G235E, G235F, G235H, G235I, G235L, G235M, G235N, G235P, G235S,G235V, S237C, S237G, S237M, S237N, S237W, S237Y, Y244C, Y244E, Y244M,L249H, L249K, L249Q, L249R, L249W, L249Y, S251L, S251N, S251R, S251W,N252P, N252C, G253D, G253W, F254I, F254L, F254M, F254Y, Q256E, Q256R,N260*, N260A, N260C, N260E, N260I, N260K, N260L, N260M, N260Q, N260R,N260V, N260W, N260Y, E261*, E261A, E261D, E261R, E261W, Q262*, Q262F,Q262H, Q262W, Q262Y, M263K, M263L, M263Q, D264*, D264C, D264E, Y265F,N267S, N267T, W268C, W268E, W268M, W268R, Y270F, A271D, A271G, H272D,H272I, H272M, H272P, H272V, H272W, N273W, K274R, K274A, K274H, F276A,F276C, F276K, F276N, F276G, F276L, F276M, F276P, F276S, F276V, F276W,I278A, I278K, I278N, I278Q, I278V, S279C, S279D, S279E, S279G, S279N,D280C, D280E, Y281*, Y281A, Y281C, Y281H, Y281K, Y281N, Y281P, Y281R,Y282E, Y282N, H283I, A284I, A284L, A284N, A284P, A284T, A284V, T287N,S288D, S288K, S288N, V290I, K291L, K291R, K291V, T296C, E300A, E300D,H301C, H301N, H301R, T303A, T303C, T303G, T303K, T303Q, T303R, T303W,D304C, D304M, L305M, L305N, S306C, K308A, K308D, K308G, K308I, K308L,K308Q, K308S, K308T, K308V, K308Y, K309A, K309C, K309D, K309H, K309L,K309M, K309N, K309Q, K309S, K309T, K309I, K312A, K312L, K312M, K312N,K312Q, K312S, K312W, E314I, E314L, E314V, L315I, L315V, R319A, andN323R, wherein the positions correspond to the positions of SEQ ID NO: 1(numbering according to SEQ ID NO: 1), wherein the variant has asequence identity to the polypeptide shown in SEQ ID NO: 1 or to thepolypeptide sequence shown in SEQ ID NO: 15 of at least 65%, at least70%, at least 75%, at least 80%, at least 85%, at least 90%, at least91%, at least 92%, at least 93%, at least 94%, at least 95%, at least96%, at least 97%, at least 98% or at least 99% but less than 100%,wherein the variant has hexosaminidase e.g. beta-1,6N-acetylglucosaminidase activity and wherein the variant has improvedstability, measured as having an improvement factor e.g. RAR or HIF>1.0,compared to the polypeptide sequence shown in SEQ ID NO: 1 or comparedto the polypeptide sequence shown in SEQ ID NO: 15.

One preferred aspect of the invention relates to a dispersin variant,which compared to a dispersin with SEQ ID NO: 1, comprises one or morealteration e.g. substitution selected from the group consisting of: D2V,Q3F, Q3I, S12A, H15Y, T17W, T17E, V18L, K22M, S23I, S23C, S23T, S23L,S23V, S23E, V25R, Y30L, A49W, A49Y, S56T, N59D, N59C, N59E, N59R, N59F,N59W, N59V, N62C, N62D, T63C, N68Q, S72D, S72E, I74L, S77A, I82V, L90F,E99Q, L100S, T114S, T114C, Y123W, Y124I, Y124M, Y124R, Y124V, Y124Q,Y124T, Y124K, D125R, D125C, D125G, D125K, D125Q, Q135M, D138R, D138K,D138Q, L170D, L170K, L170S, L170H, D171E, D171K, D171Y, D171M, D171Q,D171L, I173C, D174W, D174H, D174M, D174N, F175Y, Q178K, I179T, S181T,S181F, S181Q, S181G, S181N, S181C, E185R, E185M, E185V, S186K, S186M,S186R, S186H, K187G, Y188P, E189V, S199C, S199L, A203G, A203E, A203V,N204L, N204Y, N204V, L205I, D207N, D207S, D207C, D207G, S210T, Q215R,T218Q, N227T, N227K, E232D, G235W, S237W, Y244M, Y244C, N252P, N252C,Q256E, N260Q, Q262H, M263Q, D264E, Y265F, N267T, N267S, Y270F, H272M,H272P, H272I, H272V, N273W, K274H, F276A, F276N, F276K, F276C, I278V,S279N, S279D, S279G, D280E, D280C, Y281P, Y282N, H283I, A284T, A284L,A284I, S288K, V290I, K291R, T296C, T303Q, D304M, D304C, L305M, L305N,S306C, K308D, K308A, K308V, K308Q, K308S, K308Y, K308G, K308L, K308T,K308I, K309C, K309L, K309D, K309Q, K309N, K309T, K309A, K309S, K309M,K309H, K312A, K312Q, K312S, K312W, K312L, K312N, E314L, E314V, E314I,L315I, R319A and N323R, wherein the positions correspond to thepositions of SEQ ID NO: 1 (numbering according to SEQ ID NO: 1), whereinthe variant has a sequence identity to the polypeptide shown in SEQ IDNO: 1 the polypeptide sequence shown in SEQ ID NO: 15 of at least 65%,at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 91%, at least 92%, at least 93%, at least 94%, at least 95%, atleast 96%, at least 97%, at least 98% or at least 99% but less than100%, wherein the variant has hexosaminidase e.g. beta-1,6N-acetylglucosaminidase activity and wherein the variant has improvedstability, measured as having an improvement factor e.g. RAR or HIF>1.0,compared to the polypeptide sequence shown in SEQ ID NO: 1 or comparedto the polypeptide sequence shown in SEQ ID NO: 15.

In some preferred aspects, a dispersin variant of the invention comprisean alteration at one or more positions selected from the groupconsisting of 2, 3, 12, 15, 17, 18, 19, 22, 23, 24, 25, 26, 30, 32, 34,43, 44, 45, 49, 52, 54, 56, 57, 58, 59, 60, 62, 63, 67, 68, 71, 72, 74,77, 79, 80, 81, 82, 90, 99, 100, 103, 104, 105, 106, 107, 110, 111, 113,114, 116, 117, 118, 119, 120, 122, 123, 124, 125, 126, 127, 128, 131,135, 138, 139, 140, 142, 145, 147, 148, 149, 150, 151, 152, 163, 164,167, 168, 170, 171, 173, 174, 175, 177, 178, 179, 181, 185, 186, 187,188, 189, 199, 200, 203, 204, 205, 207, 208, 210, 215, 217, 218, 221,222, 224, 225, 227, 230, 232, 233, 234, 235, 237, 244, 249, 251, 252,253, 254, 256, 260, 261, 262, 263, 264, 265, 267, 268, 270, 271, 272,273, 274, 276, 278, 279, 280, 281, 282, 283, 284, 287, 288, 290, 291,296, 300, 301, 303, 304, 305, 306, 308, 309, 312, 314, 315, 319, 321 and323, wherein the variant has at least 60%, at least 65%, at least 70%,at least 75%, at least 80%, at least 85%, at least 90%, at least 95%,such as at least 96%, at least 97%, at least 98%, or at least 99%, butless than 100%, sequence identity to the polypeptide shown in SEQ ID NO:1, wherein each position corresponds to the position of the polypeptideshown in SEQ ID NO: 1 (numbering according to SEQ ID NO: 1) and whereinthe variant has at least one improved property compared to the parentdispersin e.g. a dispersin comprising the polypeptide having the aminoacid sequence shown in SEQ ID NO: 1, preferably the improved property isimproved stability, wherein stability is tested as described in example2a, 2b or 3.

In some preferred aspects, a dispersin variant of the invention comprisean alteration at one or more positions selected from the groupconsisting of 2, 3, 12, 15, 17, 18, 22, 23, 24, 25, 30, 49, 56, 57, 59,62, 63, 68, 72, 74, 77, 82, 90, 99, 100, 106, 114, 123, 124, 125, 135,138, 140, 163, 167, 170, 171, 173, 174, 175, 178, 179, 181, 185, 186,187, 188, 189, 199, 203, 204, 205, 207, 210, 215, 218, 221, 225, 227,232, 235, 237, 244, 252, 256, 260, 262, 263, 264, 265, 267, 270, 272,273, 274, 276, 278, 279, 280, 281, 282, 283, 284, 288, 290, 291, 296,303, 304, 305, 306, 308, 309, 312, 314, 315, 319, 321, 322 and 323,wherein the variant has at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 95%, suchas at least 96%, at least 97%, at least 98%, or at least 99%, but lessthan 100%, sequence identity to the polypeptide shown in SEQ ID NO: 1,wherein each position corresponds to the position of the polypeptideshown in SEQ ID NO: 1 (numbering according to SEQ ID NO: 1) and whereinthe variant has at least one improved property compared to the parentdispersin e.g. a dispersin comprising the polypeptide having the aminoacid sequence shown in SEQ ID NO: 1, preferably the improved property isimproved stability, wherein stability is tested as described in example2a, 2b or 3.

In some preferred aspects, a dispersin variant of the invention comprisean alteration at one or more positions selected from the groupconsisting of: 31, 33, 36, 73, 75, 94, 141, 144, 241, 277, 294, 297,299, wherein the variant has at least 60%, at least 65%, at least 70%,at least 75%, at least 80%, at least 85%, at least 90%, at least 95%,such as at least 96%, at least 97%, at least 98%, or at least 99%, butless than 100%, sequence identity to the polypeptide shown in SEQ ID NO:1, wherein each position corresponds to the position of the polypeptideshown in SEQ ID NO: 1 (numbering according to SEQ ID NO: 1) and whereinthe variant has at least one improved property compared to the parentdispersin e.g. a dispersin comprising the polypeptide having the aminoacid sequence shown in SEQ ID NO: 1, preferably the improved property isimproved stability, wherein stability is tested as described in example2a, 2b or 3.

In some preferred aspects, a dispersin variant of the invention compriseone or more alteration selected from the group consisting of: D2A, D2L,D2N, D2R, D2V, D2W, Q3F, Q3I, Q3L, Q3M, Q3P, Q3V, Q3Y, Q3T, S12A, H15F,H15Y, T17W, T17C, T17E, T17F, T17M, T17R, T17V, V18L, E19D, E19K, E19N,E19P, K22A, K22M, K22V, S23A, S23C, S23E, S23I, S23L, S23R, S23T, S23V,L24I, V25R, D26M, Y30*, Y30A, Y30D, Y30L, Y30M, Y30N, Y30R, Y30T, Y30V,G32L, G32M, G32R, N34D, N43*, N43H, N43L, E44*, N45D, N45L, N45V, A49W,A49Y, Y52*, Y52M, G54L, G54M, G54N, S56T, S56W, S57W, E58N, N59A, N59C,N59D, N59E, N59F, N59M, N59R, N59V, N59W, T60V, N62C, N62D, N62H, N62Q,N62W, T63C, T63D, T63L, T63N, T63R, T63V, K67A, K67L, N68L, N68Q, L71H,L71N, L71R, L71V, L71W, S72*, S72C, S72D, S72E, S72F, S72G, S72I, S72M,S72N, S72R, S72T, S72Y, I74L, S77A, D79V, K80*, K80E, K80H, K80L, K80N,K80Q, K80R, K80V, K80W, D81A, D81G, D81L, D81N, D81R, D81S, D81T, D81V,D81W, I82V, L90F, E99Q, E99R, L100S, K103A, K103R, K103V, K104N, K104W,D105N, V106A, V106D, V106E, V106H, V106K, V106L, V106M, V106N, V106Q,V106R, V106W, V106Y, K107A, K107C, K107L, K107M, K107T, K107V, K107W,N110M, N110R, N110V, D111A, D111E, D111M, D111N, D111Q, D111R, D111V,D111W, V113T, T114C, T114S, Y116D, Y116N, Y116R, S117*, S117D, S117H,S117N, S117P, E118*, E118A, E118D, E118G, E118L, E119G, E119W, T120I,T120L, T120M, T120V, T120W, D122*, D122H, D122R, Y123W, Y124C, Y124H,Y124I, Y124K, Y124L, Y124M, Y124N, Y124Q, Y124R, Y124T, Y124V, Y124W,D125C, D125G, D125H, D125K, D125Q, D125R, N126V, R127D, R127H, R127K,R127L, R127M, R127Q, R127W, V128A, V128C, V128D, V128L, V128T, D131V,Q135*, Q135A, Q135D, Q135E, Q135K, Q135M, Q135Y, D138K, D138L, D138M,D138N, D138Q, D138R, D138S, D138V, D138W, E139W, V140I, D142R, D142W,Y145*, Y145H, Y145L, Y145N, Y145V, P147A, P147C, P147D, P147F, P147G,P147L, P147M, P147R, P147S, P147T, P147V, K148A, K148D, K148L, K148V,F149L, F149M, F149N, E150A, E150D, E150H, E150K, E150L, E150M, E150N,E150R, E150V, E150W, E150Y, G151A, G151C, G151D, G151L, G151N, G151P,G151S, G151W, K152D, K152L, K152R, S163P, G164D, G164E, G164H, G164S,G164V, V167A, V167D, V167E, V167L, V167P, V167Q, V167R, V167W, H168N,L170A, L170D, L170E, L170F, L170H, L170K, L170M, L170N, L170P, L170Q,L170R, L170S, L170V, L170W, L170Y, D171A, D171C, D171E, D171K, D171L,D171M, D171Q, D171R, D171V, D171W, D171Y, I173C, D174H, D174M, D174N,D174V, D174W, F175Y, N177M, Q178*, Q178A, Q178K, Q178R, Q178W, I179T,S181C, S181D, S181F, S181G, S181K, S181N, S181P, S181Q, S181T, S181V,S181W, E185A, E185M, E185R, E185V, E185W, S186D, S186E, S186H, S186I,S186K, S186L, S186M, S186N, S186Q, S186R, S186V, S186W, K187C, K187D,K187G, K187R, K187S, K187V, K187W, Y188P, E189L, E189V, E189W, S199C,S199L, S199M, S199Y, E200D, E200F, E200K, E200L, E200M, E200N, E200R,E200W, A203C, A203D, A203E, A203G, A203L, A203M, A203P, A203Q, A203R,A203S, A203T, A203V, A203W, N204L, N204M, N204V, N204W, N204Y, L205I,D207A, D207C, D207E, D207G, D207K, D207N, D207Q, D207R, D207S, D207V,D207W, S208A, S208C, S208D, S208G, S208L, S208Q, S208T, S208V, S208W,S210T, Q215K, Q215R, Q215M, Q215L, Q215*, S217V, T218A, T218L, T218Q,T218R, T218V, S221N, G222D, E224A, E224P, S225G, N227A, N227Q, N227R,N227S, N227T, N227K, D230*, D230N, D230R, D230T, D230W, E232D, E232V,N233D, N233E, N233H, N233Q, N233R, N233W, W234R, G235W, G235A, G235E,G235F, G235H, G235I, G235L, G235M, G235N, G235P, G235S, G235V, S237C,S237G, S237M, S237N, S237W, S237Y, Y244C, Y244E, Y244M, L249H, L249K,L249Q, L249R, L249W, L249Y, S251A, S251L, S251N, S251R, S251W, N252P,N252C, G253D, G253W, F254I, F254L, F254M, F254Y, Q256D, Q256E, Q256R,N260*, N260A, N260C, N260E, N260I, N260K, N260L, N260M, N260Q, N260R,N260T, N260V, N260W, N260Y, E261*, E261A, E261D, E261R, E261W, Q262*,Q262F, Q262H, Q262W, Q262Y, M263K, M263L, M263Q, D264*, D264C, D264E,D264N, Y265F, N267S, N267T, W268C, W268E, W268M, W268R, Y270F, A271D,A271G, H272D, H272I, H272M, H272P, H272V, H272W, N273W, K274R, K274A,K274H, F276A, F276C, F276K, F276N, F276G, F276L, F276M, F276P, F276S,F276V, F276W, I278A, I278K, I278N, I278Q, I278V, S279C, S279D, S279E,S279G, S279N, D280C, D280E, Y281*, Y281A, Y281C, Y281H, Y281K, Y281N,Y281P, Y281R, Y282E, Y282N, H283I, A284I, A284L, A284N, A284P, A284T,A284V, T287N, S288P, S288D, S288K, S288N, V290I, K291L, K291R, K291V,T296C, E300A, E300D, H301C, H301N, H301R, T303A, T303C, T303G, T303K,T303Q, T303R, T303W, D304C, D304M, L305M, L305N, S306C, K308A, K308D,K308E, K308G, K308I, K308L, K308Q, K308S, K308T, K308V, K308Y, K309A,K309C, K309D, K309E, K309G, K309H, K309L, K309M, K309N, K309Q, K309S,K309T, K309I, K312A, K312E, K312L, K312M, K312N, K312Q, K312S, K312W,E314I, E314L, E314V, L315I, L315V, R319A, Y321F and N323R, wherein thevariant has at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%,sequence identity to the polypeptide shown in SEQ ID NO: 1, wherein eachposition corresponds to the position of the polypeptide shown in SEQ IDNO: 1 (numbering according to SEQ ID NO: 1) and wherein the variant hasat least one improved property compared to the parent dispersin e.g. adispersin comprising the polypeptide having the amino acid sequenceshown in SEQ ID NO: 1, preferably the improved property is improvedstability, wherein stability is tested as described in example 2a, 2b or3.

In some preferred aspects, a dispersin variant of the invention compriseone or more alteration selected from the group consisting of D2V, Q3F,Q3I, S12A, H15Y, T17W, T17E, V18L, K22M, S23I, S23C, S23T, S23L, S23V,S23E, V25R, Y30L, A49W, A49Y, S56T, N59D, N59C, N59E, N59R, N59F, N59W,N59V, N62C, N62D, T63C, N68Q, S72D, S72E, I74L, S77A, I82V, L90F, E99Q,L100S, T114S, T114C, Y123W, Y124I, Y124M, Y124R, Y124V, Y124Q, Y124T,Y124K, D125R, D125C, D125G, D125K, D125Q, Q135M, D138R, D138K, D138Q,L170D, L170K, L170S, L170H, D171E, D171K, D171Y, D171M, D171Q, D171L,I173C, D174W, D174H, D174M, D174N, F175Y, Q178K, I179T, S181T, S181F,S181Q, S181G, S181N, S181C, E185R, E185M, E185V, S186K, S186M, S186R,S186H, K187G, Y188P, E189V, S199C, S199L, A203G, A203E, A203V, N204L,N204Y, N204V, L205I, D207N, D207S, D207C, D207G, S210T, Q215R, T218K,T218Q, N227T, N227K, E232D, G235W, S237W, Y244M, Y244C, N252P, N252C,Q256E, N260Q, Q262H, M263Q, D264E, Y265F, N267T, N267S, Y270F, H272M,H272P, H272I, H272V, N273W, K274H, F276A, F276N, F276K, F276C, I278V,S279N, S279D, S279G, D280E, D280C, Y281P, Y282N, H283I, A284T, A284L,A284I, S288K, V290I, K291R, T296C, T303Q, D304M, D304C, L305M, L305N,S306C, K308D, K308A, K308V, K308Q, K308S, K308Y, K308G, K308L, K308T,K308I, K309C, K309L, K309D, K309Q, K309N, K309T, K309A, K309S, K309M,K309H, K312A, K312Q, K312S, K312W, K312L, K312N, E314L, E314V, E314I,L315I, R319A and N323R, wherein the variant has at least 60%, at least65%, at least 70%, at least 75%, at least 80%, at least 85%, at least90%, at least 95%, such as at least 96%, at least 97%, at least 98%, orat least 99%, but less than 100%, sequence identity to the polypeptideshown in SEQ ID NO: 1, wherein each position corresponds to the positionof the polypeptide shown in SEQ ID NO: 1 (numbering according to SEQ IDNO: 1), and wherein the variant has at least one improved propertycompared to the parent dispersin e.g. a dispersin comprising thepolypeptide having the amino acid sequence shown in SEQ ID NO: 1,preferably the improved property is improved stability, whereinstability is tested as described in example 2a, 2b or 3.

In some preferred aspects, a dispersin variant of the invention comprisean alteration at one or more positions selected from the groupconsisting of 2, 3, 12, 15, 17, 18, 19, 22, 23, 24, 25, 26, 30, 32, 34,43, 44, 45, 49, 52, 54, 56, 57, 58, 59, 60, 62, 63, 67, 68, 71, 72, 74,77, 79, 80, 81, 82, 90, 99, 100, 103, 104, 105, 106, 107, 110, 111, 113,114, 116, 117, 118, 119, 120, 122, 123, 124, 125, 126, 127, 128, 131,135, 138, 139, 140, 142, 145, 147, 148, 149, 150, 151, 152, 163, 164,167, 168, 170, 171, 173, 174, 175, 177, 178, 179, 181, 185, 186, 187,188, 189, 199, 200, 203, 204, 205, 207, 208, 210, 215, 217, 218, 221,222, 224, 225, 227, 230, 232, 233, 234, 235, 237, 244, 249, 251, 252,253, 254, 256, 260, 261, 262, 263, 264, 265, 267, 268, 270, 271, 272,273, 274, 276, 278, 279, 280, 281, 282, 283, 284, 287, 288, 290, 291,296, 300, 301, 303, 304, 305, 306, 308, 309, 312, 314, 315, 319, 321 and323, wherein each position corresponds to the position of thepolypeptide of SEQ ID NO: 1, wherein each alteration provides adispersin variant having an increase in stability measured as half-lifeimprovement factor, HIF or residual activity ratio, RAR, of at least1.05, such as 1.08, such as 1.1, such as 1.15, such as 1.2, such as1.25, such as 1.3, such as 1.4, such as 1.5, such as 1.6, such as 1.7,such as 1.8, such as 1.9, such as 2, such as 3, such as 4, such as 5 orsuch as at least 10 compared to the parent dispersin e.g. a dispersincomprising the polypeptide having the amino acid sequence shown in SEQID NO: 1, wherein the variant has at least 60%, at least 65%, at least70%, at least 75%, at least 80%, at least 85%, at least 90%, at least95%, such as at least 96%, at least 97%, at least 98%, or at least 99%,but less than 100% sequence identity to the polypeptide shown in SEQ IDNO: 1, wherein each position corresponds to the position of thepolypeptide shown in SEQ ID NO: 1 (numbering according to SEQ ID NO: 1).

In some preferred aspects, a dispersin variant of the invention comprisean alteration at one or more positions selected from the groupconsisting of: 2, 3, 12, 15, 17, 18, 22, 23, 24, 25, 30, 49, 56, 57, 59,62, 63, 68, 72, 74, 77, 82, 90, 99, 100, 106, 114, 123, 124, 125, 135,138, 140, 163, 167, 170, 171, 173, 174, 175, 178, 179, 181, 185, 186,187, 188, 189, 199, 203, 204, 205, 207, 210, 215, 218, 221, 225, 227,232, 235, 237, 244, 252, 256, 260, 262, 263, 264, 265, 267, 270, 272,273, 274, 276, 278, 279, 280, 281, 282, 283, 284, 288, 290, 291, 296,303, 304, 305, 306, 308, 309, 312, 314, 315, 319, 321, 322 and 323,wherein each substitution provides a dispersin variant having anincrease in stability measured as half-life improvement factor, HIF orresidual activity ratio, RAR, of at least 1.05, such as 1.08, such as1.1, such as 1.15, such as 1.2, such as 1.25, such as 1.3, such as 1.4,such as 1.5, such as 1.6, such as 1.7, such as 1.8, such as 1.9, such as2, such as 3, such as 4, such as 5 or such as at least 10 compared tothe parent dispersin e.g. a dispersin comprising the polypeptide havingthe amino acid sequence shown in SEQ ID NO: 1, wherein the variant hasat least 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100% sequence identityto the polypeptide shown in SEQ ID NO: 1, wherein each positioncorresponds to the position of the polypeptide shown in SEQ ID NO: 1(numbering according to SEQ ID NO: 1) and preferably wherein the varianthas hexosaminidase e.g. beta-1,6 N-acetylglucosaminidase activity.

In some preferred aspects, a dispersin variant of the invention comprisean alteration at one or more positions selected from the groupconsisting of: 31, 33, 36, 73, 75, 94, 141, 144, 241, 277, 294, 297,299, wherein each substitution provides a dispersin variant having anincrease in stability measured as half-life improvement factor, HIF orresidual activity ratio, RAR, of at least 1.05, such as 1.08, such as1.1, such as 1.15, such as 1.2, such as 1.25, such as 1.3, such as 1.4,such as 1.5, such as 1.6, such as 1.7, such as 1.8, such as 1.9, such as2, such as 3, such as 4, such as 5 or such as at least 10 compared tothe parent dispersin e.g. a dispersin comprising the polypeptide havingthe amino acid sequence shown in SEQ ID NO: 1, wherein the variant hasat least 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100% sequence identityto the polypeptide shown in SEQ ID NO: 1, wherein each positioncorresponds to the position of the polypeptide shown in SEQ ID NO: 1(numbering according to SEQ ID NO: 1) and preferably wherein the varianthas hexosaminidase e.g. beta-1,6 N-acetylglucosaminidase activity.

In some preferred aspects, a dispersin variant of the inventioncomprises one or more alteration (compared to SEQ ID NO: 1), wherein thealteration is selected from the group consisting of: D2A, D2L, D2N, D2R,D2V, D2W, Q3F, Q3I, Q3L, Q3M, Q3P, Q3V, Q3Y, Q3T, S12A, H15F, H15Y,T17W, T17C, T17E, T17F, T17M, T17R, T17V, V18L, E19D, E19K, E19N, E19P,K22A, K22M, K22V, S23A, S23C, S23E, S23I, S23L, S23R, S23T, S23V, L24I,V25R, D26M, Y30*, Y30A, Y30D, Y30L, Y30M, Y30N, Y30R, Y30T, Y30V, G32L,G32M, G32R, N34D, N43*, N43H, N43L, E44*, N45D, N45L, N45V, A49W, A49Y,Y52*, Y52M, G54L, G54M, G54N, S56T, S56W, S57W, E58N, N59A, N59C, N59D,N59E, N59F, N59M, N59R, N59V, N59W, T60V, N62C, N62D, N62H, N62Q, N62W,T63C, T63D, T63L, T63N, T63R, T63V, K67A, K67L, N68L, N68Q, L71H, L71N,L71R, L71V, L71W, S72*, S72C, S72D, S72E, S72F, S72G, S72I, S72M, S72N,S72R, S72T, S72Y, I74L, S77A, D79V, K80*, K80E, K80H, K80L, K80N, K80Q,K80R, K80V, K80W, D81A, D81G, D81L, D81N, D81R, D81S, D81T, D81V, D81W,I82V, L90F, E99Q, E99R, L100S, K103A, K103R, K103V, K104N, K104W, D105N,V106A, V106D, V106E, V106H, V106K, V106L, V106M, V106N, V106Q, V106R,V106W, V106Y, K107A, K107C, K107L, K107M, K107T, K107V, K107W, N110M,N110R, N110V, D111A, D111E, D111M, D111N, D111Q, D111R, D111V, D111W,V113T, T114C, T114S, Y116D, Y116N, Y116R, S117*, S117D, S117H, S117N,S117P, E118*, E118A, E118D, E118G, E118L, E119G, E119W, T120I, T120L,T120M, T120V, T120W, D122*, D122H, D122R, Y123W, Y124C, Y124H, Y124I,Y124K, Y124L, Y124M, Y124N, Y124Q, Y124R, Y124T, Y124V, Y124W, D125C,D125G, D125H, D125K, D125Q, D125R, N126V, R127D, R127H, R127K, R127L,R127M, R127Q, R127W, V128A, V128C, V128D, V128L, V128T, D131V, Q135*,Q135A, Q135D, Q135E, Q135K, Q135M, Q135Y, D138K, D138L, D138M, D138N,D138Q, D138R, D138S, D138V, D138W, E139W, V140I, D142R, D142W, Y145*,Y145H, Y145L, Y145N, Y145V, P147A, P147C, P147D, P147F, P147G, P147L,P147M, P147R, P147S, P147T, P147V, K148A, K148D, K148L, K148V, F149L,F149M, F149N, E150A, E150D, E150H, E150K, E150L, E150M, E150N, E150R,E150V, E150W, E150Y, G151A, G151C, G151D, G151L, G151N, G151P, G151S,G151W, K152D, K152L, K152R, S163P, G164D, G164E, G164H, G164S, G164V,V167A, V167D, V167E, V167L, V167P, V167Q, V167R, V167W, H168N, L170A,L170D, L170E, L170F, L170H, L170K, L170M, L170N, L170P, L170Q, L170R,L170S, L170V, L170W, L170Y, D171A, D171C, D171E, D171K, D171L, D171M,D171Q, D171R, D171V, D171W, D171Y, I173C, D174H, D174M, D174N, D174V,D174W, F175Y, N177M, Q178*, Q178A, Q178K, Q178R, Q178W, I179T, S181C,S181D, S181F, S181G, S181K, S181N, S181P, S181Q, S181T, S181V, S181W,E185A, E185M, E185R, E185V, E185W, S186D, S186E, S186H, S186I, S186K,S186L, S186M, S186N, S186Q, S186R, S186V, S186W, K187C, K187D, K187G,K187R, K187S, K187V, K187W, Y188P, E189L, E189V, E189W, S199C, S199L,S199M, S199Y, E200D, E200F, E200K, E200L, E200M, E200N, E200R, E200W,A203C, A203D, A203E, A203G, A203L, A203M, A203P, A203Q, A203R, A203S,A203T, A203V, A203W, N204L, N204M, N204V, N204W, N204Y, L205I, D207A,D207C, D207E, D207G, D207K, D207N, D207Q, D207R, D207S, D207V, D207W,S208A, S208C, S208D, S208G, S208L, S208Q, S208T, S208V, S208W, S210T,Q215K, Q215R, Q215M, Q215L, Q215*, S217V, T218A, T218L, T218Q, T218R,T218V, S221N, G222D, E224A, E224P, S225G, N227A, N227Q, N227R, N227S,N227T, N227K, D230*, D230N, D230R, D230T, D230W, E232D, E232V, N233D,N233E, N233H, N233Q, N233R, N233W, W234R, G235W, G235A, G235E, G235F,G235H, G235I, G235L, G235M, G235N, G235P, G235S, G235V, S237C, S237G,S237M, S237N, S237W, S237Y, Y244C, Y244E, Y244M, L249H, L249K, L249Q,L249R, L249W, L249Y, S251A, S251L, S251N, S251R, S251W, N252P, N252C,G253D, G253W, F254I, F254L, F254M, F254Y, Q256D, Q256E, Q256R, N260*,N260A, N260C, N260E, N260I, N260K, N260L, N260M, N260Q, N260R, N260T,N260V, N260W, N260Y, E261*, E261A, E261D, E261R, E261W, Q262*, Q262F,Q262H, Q262W, Q262Y, M263K, M263L, M263Q, D264*, D264C, D264E, D264N,Y265F, N267S, N267T, W268C, W268E, W268M, W268R, Y270F, A271D, A271G,H272D, H272I, H272M, H272P, H272V, H272W, N273W, K274R, K274A, K274H,F276A, F276C, F276K, F276N, F276G, F276L, F276M, F276P, F276S, F276V,F276W, I278A, I278K, I278N, I278Q, I278V, S279C, S279D, S279E, S279G,S279N, D280C, D280E, Y281*, Y281A, Y281C, Y281H, Y281K, Y281N, Y281P,Y281R, Y282E, Y282N, H283I, A284I, A284L, A284N, A284P, A284T, A284V,T287N, S288P, S288D, S288K, S288N, V290I, K291L, K291R, K291V, T296C,E300A, E300D, H301C, H301N, H301R, T303A, T303C, T303G, T303K, T303Q,T303R, T303W, D304C, D304M, L305M, L305N, S306C, K308A, K308D, K308E,K308G, K308I, K308L, K308Q, K308S, K308T, K308V, K308Y, K309A, K309C,K309D, K309E, K309G, K309H, K309L, K309M, K309N, K309Q, K309S, K309T,K309I, K312A, K312E, K312L, K312M, K312N, K312Q, K312S, K312W, E314I,E314L, E314V, L315I, L315V, R319A, Y321F and N323R, wherein eachsubstitution provides a dispersin variant having an increase instability measured as half-life improvement factor, HIF, or residualactivity, RAR, of at least 1.05, such as 1.08, such as 1.1, such as1.15, such as 1.2, such as 1.25, such as 1.3, such as 1.4, such as 1.5,such as 1.6, such as 1.7, such as 1.8, such as 1.9, such as 2, such as3, such as 4, such as 5 or such as at least 10 compared to the parentdispersin e.g. a dispersin comprising the polypeptide having the aminoacid sequence shown in SEQ ID NO: 1, wherein the variant has at least60%, at least 65%, at least 70%, at least 75%, at least 80%, at least85%, at least 90%, at least 95%, such as at least 96%, at least 97%, atleast 98%, or at least 99%, but less than 100% sequence identity to thepolypeptide shown in SEQ ID NO: 1, wherein each position corresponds tothe position of the polypeptide shown in SEQ ID NO: 1 (numberingaccording to SEQ ID NO: 1).

In some preferred aspects, a dispersin variant of the inventioncomprises one or more substitutions (compared to SEQ ID NO: 1), whereinthe substitutions are selected from the group consisting of: D2V, Q3F,Q3I, S12A, H15Y, T17W, T17E, V18L, K22M, S23I, S23C, S23T, S23L, S23V,S23E, V25R, Y30L, A49W, A49Y, S56T, N59D, N59C, N59E, N59R, N59F, N59W,N59V, N62C, N62D, T63C, N68Q, S72D, S72E, I74L, S77A, I82V, L90F, E99Q,L100S, T114S, T114C, Y123W, Y124I, Y124M, Y124R, Y124V, Y124Q, Y124T,Y124K, D125R, D125C, D125G, D125K, D125Q, Q135M, D138R, D138K, D138Q,L170D, L170K, L170S, L170H, D171E, D171K, D171Y, D171M, D171Q, D171L,I173C, D174W, D174H, D174M, D174N, F175Y, Q178K, I179T, S181T, S181F,S181Q, S181G, S181N, S181C, E185R, E185M, E185V, S186K, S186M, S186R,S186H, K187G, Y188P, E189V, S199C, S199L, A203G, A203E, A203V, N204L,N204Y, N204V, L205I, D207N, D207S, D207C, D207G, S210T, Q215R, T218Q,S225G, N227T, N227K, E232D, G235W, S237W, Y244M, Y244C, N252P, N252C,Q256E, N260Q, Q262H, M263Q, D264E, Y265F, N267T, N267S, Y270F, H272M,H272P, H272I, H272V, N273W, K274H, F276A, F276N, F276K, F276C, I278V,S279N, S279D, S279G, D280E, D280C, Y281P, Y282N, H283I, A284T, A284L,A284I, S288K, V290I, K291R, T296C, T303Q, D304M, D304C, L305M, L305N,S306C, K308D, K308A, K308V, K308Q, K308S, K308Y, K308G, K308L, K308T,K308I, K309C, K309L, K309D, K309Q, K309N, K309T, K309A, K309S, K309M,K309H, K312A, K312Q, K312S, K312W, K312L, K312N, E314L, E314V, E314I,L315I, R319A and N323R, wherein each substitution provides a dispersinvariant having an increase in stability measured as half-lifeimprovement factor, HIF or residual activity ratio, RAR, of at least1.05, such as 1.08, such as 1.1, such as 1.15, such as 1.2, such as1.25, such as 1.3, such as 1.4, such as 1.5, such as 1.6, such as 1.7,such as 1.8, such as 1.9, such as 2, such as 3, such as 4, such as 5 orsuch as at least 10 compared to the parent dispersin e.g. a dispersincomprising the polypeptide having the amino acid sequence shown in SEQID NO: 1, wherein the variant has at least 60%, at least 65%, at least70%, at least 75%, at least 80%, at least 85%, at least 90%, at least95%, such as at least 96%, at least 97%, at least 98%, or at least 99%,but less than 100% sequence identity to the polypeptide shown in SEQ IDNO: 1, wherein each position corresponds to the position of thepolypeptide shown in SEQ ID NO: 1 (numbering according to SEQ ID NO: 1).

In some preferred embodiment of the invention, the dispersin variant ofthe invention comprises one or more of the following substitutions: Q3F,Q3I, H15Y, T17W, A49W, N59E, V140I, S163P, S186R, D207N, Q215K, T218Q,S225G, N227T, E232D, G235W, S237W, N252P, N260Q, N267T, H272V, H272P,F276A, S279D, Y281P, S288P, K308Q, K308E, K309E, K312Q, K312E, whereinthe variant has at least 60%, at least 65%, at least 70%, at least 75%,at least 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%sequence identity to the polypeptide shown in SEQ ID NO: 1, wherein eachposition corresponds to the position of the polypeptide shown in SEQ IDNO: 1 (numbering according to SEQ ID NO: 1).

The variants of the invention preferably comprise one or more of theconservative motifs GXDE (SEQ ID NO:8),[EQ][NRSHA][YVFL][AGSTC][IVLF][EAQYN][SN] (SEQ IDNO:9),[VIM][LIV]G[GAV]DE[VI][PSA] SEQ ID NO: 10), WND[SQR][IVL][TLVM](SEQ ID NO: 11), QSTL (SEQ ID NO: 12), NKFFY (SEQ ID NO: 13) or NLD[DR]S(SEQ ID NO: 14), which are shared among dispersins of the Terribacillusclade as described below. As explained in “Definitions” a cladecomprises a group of polypeptides clustered together based on homologousfeatures traced to a common ancestor. Polypeptides forming a group e.g.a clade as shown in a phylogenetic tree often share common propertiesand are more closely related than other polypeptides not in the clade.

One preferred aspect of the invention relates to a variant of adispersin parent, wherein the variant comprises one or more motifs GXDE(SEQ ID NO:8), [EQ][NRSHA][YVFL][AGSTC][IVLF][EAQYN][SN] (SEQ IDNO:9),[VIM][LIV]G[GAV]DE[VI][PSA] (SEQ ID NO: 10), WND[SQR][IVL][TLVM](SEQ ID NO: 11), QSTL (SEQ ID NO: 12), NKFFY (SEQ ID NO: 13) or NLD[DR]S(SEQ ID NO: 14), and wherein the variant comprise an alteration at oneor more positions selected from the list consisting of positions: 2, 3,12, 15, 17, 18, 19, 22, 23, 24, 25, 26, 30, 32, 34, 43, 44, 45, 49, 52,54, 56, 57, 58, 59, 60, 62, 63, 67, 68, 71, 72, 74, 77, 79, 80, 81, 82,90, 99, 100, 103, 104, 105, 106, 107, 110, 111, 113, 114, 116, 117, 118,119, 120, 122, 123, 124, 125, 126, 127, 128, 131, 135, 138, 139, 140,142, 145, 147, 148, 149, 150, 151, 152, 163, 164, 167, 168, 170, 171,173, 174, 175, 177, 178, 179, 181, 185, 186, 187, 188, 189, 199, 200,203, 204, 205, 207, 208, 210, 215, 217, 218, 221, 222, 224, 225, 227,230, 232, 233, 234, 235, 237, 244, 249, 251, 252, 253, 254, 256, 260,261, 262, 263, 264, 265, 267, 268, 270, 271, 272, 273, 274, 276, 278,279, 280, 281, 282, 283, 284, 287, 288, 290, 291, 296, 300, 301, 303,304, 305, 306, 308, 309, 312, 314, 315, 319, 321 and 323, wherein eachposition corresponds to the position of the polypeptide of SEQ ID NO: 1,wherein each alteration provides a dispersin variant with at least oneimproved property compared to the polypeptide shown in SEQ ID NO: 1,wherein the variant has at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 95%, suchas at least 96%, at least 97%, at least 98%, or at least 99%, but lessthan 100% sequence identity to the polypeptide shown in SEQ ID NO: 1 andwherein each position corresponds to the position of the polypeptideshown in SEQ ID NO: 1 (numbering according to SEQ ID NO: 1) andpreferably wherein the variant has hexosaminidase e.g. beta-1,6N-acetylglucosaminidase activity.

one preferred aspect of the invention relates to a variant of adispersin parent, wherein the variant comprises one or more motifs GXDE(SEQ ID NO:8), [EQ][NRSHA][YVFL][AGSTC][IVLF][EAQYN][SN] (SEQ IDNO:9),[VIM][LIV]G[GAV]DE[VI][PSA] SEQ ID NO: 10), [WND[SQR][IVL][TLVM](SEQ ID NO: 11), QSTL (SEQ ID NO: 12), NKFFY (SEQ ID NO: 13) or NLD[DR]S(SEQ ID NO: 14), and wherein the variant comprise an alteration at oneor more positions selected from the list consisting of positions: 2, 3,12, 15, 17, 18, 22, 23, 24, 25, 30, 49, 56, 57, 59, 62, 63, 68, 72, 74,77, 82, 90, 99, 100, 106, 114, 123, 124, 125, 135, 138, 140, 163, 167,170, 171, 173, 174, 175, 178, 179, 181, 185, 186, 187, 188, 189, 199,203, 204, 205, 207, 210, 215, 218, 221, 225, 227, 232, 235, 237,244,252, 256, 260, 262, 263, 264, 265, 267, 270, 272, 273, 274, 276, 278,279, 280, 281, 282, 283, 284, 288, 290, 291, 296, 303, 304, 305, 306,308, 309, 312, 314, 315, 319, 321, 322 and 323, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereineach alteration provides a dispersin variant with at least one improvedproperty compared to the polypeptide shown in SEQ ID NO: 1, wherein thevariant has at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, at least 90%, at least 95%, such as at least96%, at least 97%, at least 98%, or at least 99%, but less than 100%sequence identity to the polypeptide shown in SEQ ID NO: 1 and whereineach position corresponds to the position of the polypeptide shown inSEQ ID NO: 1 (numbering according to SEQ ID NO: 1) and preferablywherein the variant has hexosaminidase e.g. beta-1,6N-acetylglucosaminidase activity.

one preferred aspect of the invention relates to a variant of adispersin parent, wherein the variant comprises one or more motifs GXDE(SEQ ID NO:8), [EQ][NRSHA][YVFL][AGSTC][IVLF][EAQYN][SN] (SEQ IDNO:9),[VIM][LIV]G[GAV]DE[VI][PSA] SEQ ID NO: 10), [WND[SQR][IVL][TLVM](SEQ ID NO: 11), QSTL (SEQ ID NO: 12), NKFFY (SEQ ID NO: 13) or NLD[DR]S(SEQ ID NO: 14), and wherein the variant comprise an alteration at oneor more positions selected from the list consisting of positions: 31,33, 36, 73, 75, 94, 141, 144, 241, 277, 294, 297, 299, wherein eachposition corresponds to the position of the polypeptide of SEQ ID NO: 1,wherein each alteration provides a dispersin variant with at least oneimproved property compared to the polypeptide shown in SEQ ID NO: 1,wherein the variant has at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 95%, suchas at least 96%, at least 97%, at least 98%, or at least 99%, but lessthan 100% sequence identity to the polypeptide shown in SEQ ID NO: 1 andwherein each position corresponds to the position of the polypeptideshown in SEQ ID NO: 1 (numbering according to SEQ ID NO: 1) andpreferably wherein the variant has hexosaminidase e.g. beta-1,6N-acetylglucosaminidase activity.

One preferred aspect of the invention relates to a variant of adispersin parent, wherein the variant comprises one or more motifs GXDE(SEQ ID NO:8), [EQ][NRSHA][YVFL][AGSTC][IVLF][EAQYN][SN] (SEQ IDNO:9),[VIM][LIV]G[GAV]DE[VI][PSA] SEQ ID NO: 10), [WND[SQR][IVL][TLVM](SEQ ID NO: 11), QSTL (SEQ ID NO: 12), NKFFY (SEQ ID NO: 13) or NLD[DR]S(SEQ ID NO: 14), wherein the variant comprises one or more alterations(compared to SEQ ID NO: 1), wherein the alterations are selected fromthe group consisting of: D2A, D2L, D2N, D2R, D2V, D2W, Q3F, Q3I, Q3L,Q3M, Q3P, Q3V, Q3Y, Q3T, S12A, H15F, H15Y, T17W, T17C, T17E, T17F, T17M,T17R, T17V, V18L, E19D, E19K, E19N, E19P, K22A, K22M, K22V, S23A, S23C,S23E, S23I, S23L, S23R, S23T, S23V, L24I, V25R, D26M, Y30*, Y30A, Y30D,Y30L, Y30M, Y30N, Y30R, Y30T, Y30V, G32L, G32M, G32R, N34D, N43*, N43H,N43L, E44*, N45D, N45L, N45V, A49W, A49Y, Y52*, Y52M, G54L, G54M, G54N,S56T, S56W, S57W, E58N, N59A, N59C, N59D, N59E, N59F, N59M, N59R, N59V,N59W, T60V, N62C, N62D, N62H, N62Q, N62W, T63C, T63D, T63L, T63N, T63R,T63V, K67A, K67L, N68L, N68Q, L71H, L71N, L71R, L71V, L71W, S72*, S72C,S72D, S72E, S72F, S72G, S72I, S72M, S72N, S72R, S72T, S72Y, I74L, S77A,D79V, K80*, K80E, K80H, K80L, K80N, K80Q, K80R, K80V, K80W, D81A, D81G,D81L, D81N, D81R, D81S, D81T, D81V, D81W, I82V, L90F, E99Q, E99R, L100S,K103A, K103R, K103V, K104N, K104W, D105N, V106A, V106D, V106E, V106H,V106K, V106L, V106M, V106N, V106Q, V106R, V106W, V106Y, K107A, K107C,K107L, K107M, K107T, K107V, K107W, N110M, N110R, N110V, D111A, D111E,D111M, D111N, D111Q, D111R, D111V, D111W, V113T, T114C, T114S, Y116D,Y116N, Y116R, S117*, S117D, S117H, S117N, S117P, E118*, E118A, E118D,E118G, E118L, E119G, E119W, T120I, T120L, T120M, T120V, T120W, D122*,D122H, D122R, Y123W, Y124C, Y124H, Y124I, Y124K, Y124L, Y124M, Y124N,Y124Q, Y124R, Y124T, Y124V, Y124W, D125C, D125G, D125H, D125K, D125Q,D125R, N126V, R127D, R127H, R127K, R127L, R127M, R127Q, R127W, V128A,V128C, V128D, V128L, V128T, D131V, Q135*, Q135A, Q135D, Q135E, Q135K,Q135M, Q135Y, D138K, D138L, D138M, D138N, D138Q, D138R, D138S, D138V,D138W, E139W, V140I, D142R, D142W, Y145*, Y145H, Y145L, Y145N, Y145V,P147A, P147C, P147D, P147F, P147G, P147L, P147M, P147R, P147S, P147T,P147V, K148A, K148D, K148L, K148V, F149L, F149M, F149N, E150A, E150D,E150H, E150K, E150L, E150M, E150N, E150R, E150V, E150W, E150Y, G151A,G151C, G151D, G151L, G151N, G151P, G151S, G151W, K152D, K152L, K152R,S163P, G164D, G164E, G164H, G164S, G164V, V167A, V167D, V167E, V167L,V167P, V167Q, V167R, V167W, H168N, L170A, L170D, L170E, L170F, L170H,L170K, L170M, L170N, L170P, L170Q, L170R, L170S, L170V, L170W, L170Y,D171A, D171C, D171E, D171K, D171L, D171M, D171Q, D171R, D171V, D171W,D171Y, I173C, D174H, D174M, D174N, D174V, D174W, F175Y, N177M, Q178*,Q178A, Q178K, Q178R, Q178W, I179T, S181C, S181D, S181F, S181G, S181K,S181N, S181P, S181Q, S181T, S181V, S181W, E185A, E185M, E185R, E185V,E185W, S186D, S186E, S186H, S186I, S186K, S186L, S186M, S186N, S186Q,S186R, S186V, S186W, K187C, K187D, K187G, K187R, K187S, K187V, K187W,Y188P, E189L, E189V, E189W, S199C, S199L, S199M, S199Y, E200D, E200F,E200K, E200L, E200M, E200N, E200R, E200W, A203C, A203D, A203E, A203G,A203L, A203M, A203P, A203Q, A203R, A203S, A203T, A203V, A203W, N204L,N204M, N204V, N204W, N204Y, L205I, D207A, D207C, D207E, D207G, D207K,D207N, D207Q, D207R, D207S, D207V, D207W, S208A, S208C, S208D, S208G,S208L, S208Q, S208T, S208V, S208W, S210T, Q215K, Q215R, Q215M, Q215L,Q215*, S217V, T218A, T218L, T218Q, T218R, T218V, S221N, G222D, E224A,E224P, S225G, N227A, N227Q, N227R, N227S, N227T, N227K, D230*, D230N,D230R, D230T, D230W, E232D, E232V, N233D, N233E, N233H, N233Q, N233R,N233W, W234R, G235W, G235A, G235E, G235F, G235H, G235I, G235L, G235M,G235N, G235P, G235S, G235V, S237C, S237G, S237M, S237N, S237W, S237Y,Y244C, Y244E, Y244M, L249H, L249K, L249Q, L249R, L249W, L249Y, S251A,S251L, S251N, S251R, S251W, N252P, N252C, G253D, G253W, F254I, F254L,F254M, F254Y, Q256D, Q256E, Q256R, N260*, N260A, N260C, N260E, N260I,N260K, N260L, N260M, N260Q, N260R, N260T, N260V, N260W, N260Y, E261*,E261A, E261D, E261R, E261W, Q262*, Q262F, Q262H, Q262W, Q262Y, M263K,M263L, M263Q, D264*, D264C, D264E, D264N, Y265F, N267S, N267T, W268C,W268E, W268M, W268R, Y270F, A271D, A271G, H272D, H272I, H272M, H272P,H272V, H272W, N273W, K274R, K274A, K274H, F276A, F276C, F276K, F276N,F276G, F276L, F276M, F276P, F276S, F276V, F276W, I278A, I278K, I278N,I278Q, I278V, S279C, S279D, S279E, S279G, S279N, D280C, D280E, Y281*,Y281A, Y281C, Y281H, Y281K, Y281N, Y281P, Y281R, Y282E, Y282N, H283I,A284I, A284L, A284N, A284P, A284T, A284V, T287N, S288P, S288D, S288K,S288N, V290I, K291L, K291R, K291V, T296C, E300A, E300D, H301C, H301N,H301R, T303A, T303C, T303G, T303K, T303Q, T303R, T303W, D304C, D304M,L305M, L305N, S306C, K308A, K308D, K308E, K308G, K308I, K308L, K308Q,K308S, K308T, K308V, K308Y, K309A, K309C, K309D, K309E, K309G, K309H,K309L, K309M, K309N, K309Q, K309S, K309T, K309I, K312A, K312E, K312L,K312M, K312N, K312Q, K312S, K312W, E314I, E314L, E314V, L315I, L315V,R319A, Y321F and N323R, wherein each substitution provides a dispersinvariant with at least one improved property compared to the polypeptideshown in SEQ ID NO: 1, wherein the variant has at least 60%, at least65%, at least 70%, at least 75%, at least 80%, at least 85%, at least90%, at least 95%, such as at least 96%, at least 97%, at least 98%, orat least 99%, but less than 100% sequence identity to the polypeptideshown in SEQ ID NO: 1 and wherein each position corresponds to theposition of the polypeptide shown in SEQ ID NO: 1 (numbering accordingto SEQ ID NO: 1) and preferably wherein the variant has hexosaminidasee.g. beta-1,6 N-acetylglucosaminidase activity.

One preferred aspect of the invention relates to a variant of adispersin parent, wherein the variant comprises one or more motifs GXDE(SEQ ID NO:8), [EQ][NRSHA][YVFL][AGSTC][IVLF][EAQYN][SN] (SEQ IDNO:9),[VIM][LIV]G[GAV]DE[VI][PSA] SEQ ID NO: 10), [WND[SQR][IVL][TLVM](SEQ ID NO: 11), QSTL (SEQ ID NO: 12), NKFFY (SEQ ID NO: 13) or NLD[DR]S(SEQ ID NO: 14), wherein the variant comprises one or more alterations(compared to SEQ ID NO: 1), wherein the alterations are selected fromthe group consisting of: D2A, D2L, D2N, D2R, D2V, D2W, Q3F, Q3I, Q3L,Q3M, Q3P, Q3V, Q3Y, Q3T, S12A, H15F, H15Y, T17W, T17C, T17E, T17F, T17M,T17R, T17V, V18L, E19D, E19N, E19P, K22A, K22M, K22V, S23C, S23E, S23I,S23L, S23R, S23T, S23V, V25R, D26M, Y30*, Y30D, Y30L, Y30M, Y30N, Y30R,Y30T, Y30V, G32L, G32M, G32R, N43*, N43H, N43L, E44*, N45D, N45L, N45V,A49W, A49Y, Y52*, Y52M, G54L, G54M, G54N, S56T, S56W, S57W, E58N, N59A,N59C, N59D, N59E, N59F, N59M, N59R, N59V, N59W, T60V, N62C, N62D, N62H,N62Q, N62W, T63C, T63D, T63L, T63N, T63R, T63V, K67A, K67L, N68L, N68Q,L71H, L71N, L71R, L71V, L71W, S72*, S720, S72D, S72E, S72F, S72G, S72I,S72M, S72N, S72R, S72T, S72Y, I74L, S77A, D79V, K80*, K80E, K80H, K80L,K80N, K80Q, K80V, K80W, D81A, D81G, D81L, D81R, D81S, D81T, D81V, D81W,I82V, L90F, E99Q, E99R, L100S, K103A, K103R, K104N, K104W, D105N, V106A,V106D, V106E, V106H, V106K, V106L, V106M, V106N, V106Q, V106W, V106Y,K107A, K107C, K107L, K107M, K107T, K107V, K107W, N110M, N110R, N110V,D111A, D111E, D111M, D111N, D111Q, D111R, D111V, D111W, V113T, T114C,T114S, Y116D, Y116N, Y116R, S117*, S117D, S117H, S117N, S117P, E118*,E118A, E118D, E118G, E118L, E119G, E119W, T120I, T120L, T120M, T120V,T120W, D122*, D122H, D122R, Y123W, Y124C, Y124I, Y124K, Y124L, Y124M,Y124Q, Y124R, Y124T, Y124V, Y124W, D125C, D125G, D125K, D125Q, D125R,N126V, R127D, R127H, R127K, R127L, R127M, R127Q, R127W, V128C, V128L,V128T, D131V, Q135*, Q135A, Q135D, Q135E, Q135K, Q135M, Q135Y, D138K,D138L, D138M, D138Q, D138R, D138S, D138V, D138W, E139W, D142R, D142W,Y145*, Y145H, Y145L, Y145N, Y145V, P147A, P147C, P147D, P147F, P147G,P147L, P147M, P147R, P147S, P147T, P147V, K148A, K148D, K148L, K148V,F149L, F149M, F149N, E150D, E150H, E150K, E150L, E150M, E150N, E150R,E150V, E150W, E150Y, G151A, G151C, G151D, G151L, G151N, G151P, G151S,G151W, K152D, K152L, K152R, G164D, G164E, G164H, G164S, G164V, V167D,V167E, V167L, V167P, V167Q, V167R, V167W, H168N, L170A, L170D, L170E,L170F, L170H, L170K, L170M, L170N, L170P, L170Q, L170R, L170S, L170V,L170W, L170Y, D171A, D171C, D171E, D171K, D171L, D171M, D171Q, D171R,D171V, D171W, D171Y, I173C, D174H, D174M, D174N, D174V, D174W, F175Y,N177M, Q178*, Q178A, Q178K, Q178R, Q178W, I179T, S181C, S181D, S181F,S181G, S181N, S181P, S181Q, S181T, S181V, S181W, E185M, E185R, E185V,E185W, S186D, S186E, S186H, S186I, S186K, S186L, S186M, S186N, S186Q,S186R, S186V, S186W, K187C, K187D, K187G, K187R, K187S, K187V, K187W,Y188P, E189L, E189V, E189W, S199C, S199L, S199M, S199Y, E200D, E200F,E200K, E200L, E200M, E200N, E200R, E200W, A203C, A203D, A203E, A203G,A203L, A203M, A203P, A203R, A203S, A203T, A203V, A203W, N204L, N204M,N204V, N204W, N204Y, L205I, D207A, D207C, D207E, D207G, D207N, D207Q,D207S, D207V, D207W, S208A, S208C, S208D, S208G, S208L, S208Q, S208T,S208V, S208W, S210T, Q215R, Q215M, Q215L, Q215*, S217V, T218A, T218L,T218Q, T218R, T218V, G222D, E224A, E224P, N227A, N227Q, N227R, N227S,N227T, N227K, D230*, D230R, D230T, D230W, E232D, E232V, N233H, N233Q,N233R, N233W, W234R, G235W, G235A, G235E, G235F, G235H, G235I, G235L,G235M, G235N, G235P, G235S, G235V, S237C, S237G, S237M, S237N, S237W,S237Y, Y244C, Y244E, Y244M, L249H, L249K, L249Q, L249R, L249W, L249Y,S251L, S251N, S251R, S251W, N252P, N252C, G253D, G253W, F254I, F254L,F254M, F254Y, Q256E, Q256R, N260*, N260A, N260C, N260E, N260I, N260K,N260L, N260M, N260Q, N260R, N260V, N260W, N260Y, E261*, E261A, E261D,E261R, E261W, Q262*, Q262F, Q262H, Q262W, Q262Y, M263K, M263L, M263Q,D264*, D264C, D264E, Y265F, N267S, N267T, W268C, W268E, W268M, W268R,Y270F, A271D, A271G, H272D, H272I, H272M, H272P, H272V, H272W, N273W,K274R, K274A, K274H, F276A, F276C, F276K, F276N, F276G, F276L, F276M,F276P, F276S, F276V, F276W, I278A, I278K, I278N, I278Q, I278V, S279C,S279D, S279E, S279G, S279N, D280C, D280E, Y281*, Y281A, Y281C, Y281H,Y281K, Y281N, Y281P, Y281R, Y282E, Y282N, H283I, A284I, A284L, A284N,A284P, A284T, A284V, T287N, S288D, S288K, S288N, V290I, K291L, K291R,K291V, T296C, E300A, E300D, H301C, H301N, H301R, T303A, T303C, T303G,T303K, T303Q, T303R, T303W, D304C, D304M, L305M, L305N, S306C, K308A,K308D, K308G, K308I, K308L, K308Q, K308S, K308T, K308V, K308Y, K309A,K309C, K309D, K309H, K309L, K309M, K309N, K309Q, K309S, K309T, K309I,K312A, K312L, K312M, K312N, K312Q, K312S, K312W, E314I, E314L, E314V,L315I, L315V, R319A, and N323R, wherein each substitution provides adispersin variant with at least one improved property compared to thepolypeptide shown in SEQ ID NO: 1, wherein the variant has at least 60%,at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, atleast 90%, at least 95%, such as at least 96%, at least 97%, at least98%, or at least 99%, but less than 100% sequence identity to thepolypeptide shown in SEQ ID NO: 1 and wherein each position correspondsto the position of the polypeptide shown in SEQ ID NO: 1 (numberingaccording to SEQ ID NO: 1) and preferably wherein the variant hashexosaminidase e.g. beta-1,6 N-acetylglucosaminidase activity.

One preferred aspect of the invention relates to a variant of adispersin parent, wherein the variant comprises one or moresubstitution(s) (compared to SEQ ID NO: 1), wherein the variantcomprises one or more motifs GXDE (SEQ ID NO:8),[EQ][NRSHA][YVFL][AGSTC][IVLF][EAQYN][SN] (SEQ IDNO:9),[VIM][LIV]G[GAV]DE[VI][PSA] SEQ ID NO: 10), [WND[SQR][IVL][TLVM](SEQ ID NO: 11), QSTL (SEQ ID NO: 12), NKFFY (SEQ ID NO: 13) or NLD[DR]S(SEQ ID NO: 14), wherein the substitution(s) are selected from the groupconsisting of: D2V, Q3I, Q3F, S12A, H15Y, T17W, T17E, V18L, K22M, S23I,S23C, S23T, S23L, S23V, S23E, V25R, Y30L, A49W, A49Y, S56T, N59D, N59C,N59E, N59R, N59F, N59W, N59V, N62C, N62D, T63C, N68Q, S72D, S72E, I74L,S77A, I82V, L90F, E99Q, L100S, T114S, T114C, Y123W, Y124I, Y124M, Y124R,Y124V, Y124Q, Y124T, Y124K, D125R, D125C, D125G, D125K, D125Q, Q135M,D138R, D138K, D138Q, L170D, L170K, L170S, L170H, D171E, D171K, D171Y,D171M, D171Q, D171L, I173C, D174W, D174H, D174M, D174N, F175Y, Q178K,I179T, S181T, S181F, S181Q, S181G, S181N, S181C, E185R, E185M, E185V,S186K, S186M, S186R, S186H, K187G, Y188P, E189V, S199C, S199L, A203G,A203E, A203V, N204L, N204Y, N204V, L205I, D207N, D207S, D207C, D207G,S210T, Q215R, T218Q, N227T, N227K, E232D, G235W, S237W, Y244M, Y244C,N252P, N252C, Q256E, N260Q, Q262H, M263Q, D264E, Y265F, N267T, N267S,Y270F, H272M, H272P, H272I, H272V, N273W, K274H, F276A, F276N, F276K,F276C, I278V, S279N, S279D, S279G, D280E, D280C, Y281P, Y282N, H283I,A284T, A284L, A284I, S288K, V290I, K291R, T296C, T303Q, D304M, D304C,L305M, L305N, S306C, K308D, K308A, K308V, K308Q, K308S, K308Y, K308G,K308L, K308T, K308I, K309C, K309L, K309D, K309Q, K309N, K309T, K309A,K309S, K309M, K309H, K312A, K312Q, K312S, K312W, K312L, K312N, E314L,E314V, E314I, L315I, R319A and N323R, wherein each substitution providesa dispersin variant with at least one improved property compared to thepolypeptide shown in SEQ ID NO: 1, wherein the variant has at least 60%,at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, atleast 90%, at least 95%, such as at least 96%, at least 97%, at least98%, or at least 99%, but less than 100% sequence identity to thepolypeptide shown in SEQ ID NO: 1 and wherein each position correspondsto the position of the polypeptide shown in SEQ ID NO: 1 (numberingaccording to SEQ ID NO: 1) and preferably wherein the variant hashexosaminidase e.g. beta-1,6 N-acetylglucosaminidase activity.

One preferred aspect of the invention relates to a variant of adispersin parent, wherein the variant comprises one or more motifs GXDE(SEQ ID NO:8), [EQ][NRSHA][YVFL][AGSTC][IVLF][EAQYN][SN] (SEQ IDNO:9),[VIM][LIV]G[GAV]DE[VI][PSA] SEQ ID NO: 10), [WND[SQR][IVL][TLVM](SEQ ID NO: 11), QSTL (SEQ ID NO: 12), NKFFY (SEQ ID NO: 13) or NLD[DR]S(SEQ ID NO: 14), and wherein the variant comprise an alteration at oneor more positions selected from the list consisting of positions: 2, 3,12, 15, 17, 18, 19, 22, 23, 24, 25, 26, 30, 32, 34, 43, 44, 45, 49, 52,54, 56, 57, 58, 59, 60, 62, 63, 67, 68, 71, 72, 74, 77, 79, 80, 81, 82,90, 99, 100, 103, 104, 105, 106, 107, 110, 111, 113, 114, 116, 117, 118,119, 120, 122, 123, 124, 125, 126, 127, 128, 131, 135, 138, 139, 140,142, 145, 147, 148, 149, 150, 151, 152, 163, 164, 167, 168, 170, 171,173, 174, 175, 177, 178, 179, 181, 185, 186, 187, 188, 189, 199, 200,203, 204, 205, 207, 208, 210, 215, 217, 218, 221, 222, 224, 225, 227,230, 232, 233, 234, 235, 237, 244, 249, 251, 252, 253, 254, 256, 260,261, 262, 263, 264, 265, 267, 268, 270, 271, 272, 273, 274, 276, 278,279, 280, 281, 282, 283, 284, 287, 288, 290, 291, 296, 300, 301, 303,304, 305, 306, 308, 309, 312, 314, 315, 319, 321 and 323, wherein eachposition corresponds to the position of the polypeptide of SEQ ID NO: 1,wherein each substitution provides a dispersin variant having anincrease in stability measured as half-life improvement factor, HIF orresidual activity ratio, RAR, of at least 1.05, such as 1.08, such as1.1, such as 1.15, such as 1.2, such as 1.25, such as 1.3, such as 1.4,such as 1.5, such as 1.6, such as 1.7, such as 1.8, such as 1.9, such as2, such as 3, such as 4, such as 5 or such as at least 10 compared tothe parent dispersin e.g. a dispersin comprising the polypeptide havingthe amino acid sequence shown in SEQ ID NO: 1, wherein the variant hasat least 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100% sequence identityto the polypeptide shown in SEQ ID NO: 1 and preferably wherein thevariant has hexosaminidase e.g. beta-1,6 N-acetylglucosaminidaseactivity.

One preferred aspect of the invention relates to a variant of adispersin parent, wherein the variant comprises one or more motifs GXDE(SEQ ID NO:8), [EQ][NRSHA][YVFL][AGSTC][IVLF][EAQYN][SN] (SEQ IDNO:9),[VIM][LIV]G[GAV]DE[VI][PSA] SEQ ID NO: 10), [WND[SQR][IVL][TLVM](SEQ ID NO: 11), QSTL (SEQ ID NO: 12), NKFFY (SEQ ID NO: 13) or NLD[DR]S(SEQ ID NO: 14), and wherein the variant comprise an alteration at oneor more positions selected from the list consisting of positions: 2, 3,12, 15, 17, 18, 22, 23, 24, 25, 30, 49, 56, 57, 59, 62, 63, 68, 72, 74,77, 82, 90, 99, 100, 106, 114, 123, 124, 125, 135, 138, 140, 163, 167,170, 171, 173, 174, 175, 178, 179, 181, 185, 186, 187, 188, 189, 199,203, 204, 205, 207, 210, 215, 218, 221, 225, 227, 232, 235, 237, 244,252, 256, 260, 262, 263, 264, 265, 267, 270, 272, 273, 274, 276, 278,279, 280, 281, 282, 283, 284, 288, 290, 291, 296, 303, 304, 305, 306,308, 309, 312, 314, 315, 319, 321, 322 and 323, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereineach substitution provides a dispersin variant having an increase instability measured as half-life improvement factor, HIF or residualactivity ratio, RAR, of at least 1.05, such as 1.08, such as 1.1, suchas 1.15, such as 1.2, such as 1.25, such as 1.3, such as 1.4, such as1.5, such as 1.6, such as 1.7, such as 1.8, such as 1.9, such as 2, suchas 3, such as 4, such as 5 or such as at least 10 compared to the parentdispersin e.g. a dispersin comprising the polypeptide having the aminoacid sequence shown in SEQ ID NO: 1, wherein the variant has at least60%, at least 65%, at least 70%, at least 75%, at least 80%, at least85%, at least 90%, at least 95%, such as at least 96%, at least 97%, atleast 98%, or at least 99%, but less than 100% sequence identity to thepolypeptide shown in SEQ ID NO: 1 and preferably wherein the variant hashexosaminidase e.g. beta-1,6 N-acetylglucosaminidase activity.

One preferred aspect of the invention relates to a variant of adispersin parent, wherein the variant comprises one or more motifs GXDE(SEQ ID NO:8), [EQ][NRSHA][YVFL][AGSTC][IVLF][EAQYN][SN] (SEQ IDNO:9),[VIM][LIV]G[GAV]DE[VI][PSA] SEQ ID NO: 10), [WND[SQR][IVL][TLVM](SEQ ID NO: 11), QSTL (SEQ ID NO: 12), NKFFY (SEQ ID NO: 13) or NLD[DR]S(SEQ ID NO: 14), and wherein the variant comprise an alteration at oneor more positions selected from the list consisting of positions: 31,33, 36, 73, 75, 94, 141, 144, 241, 277, 294, 297, 299, wherein eachposition corresponds to the position of the polypeptide of SEQ ID NO: 1,wherein each substitution provides a dispersin variant having anincrease in stability measured as half-life improvement factor, HIF orresidual activity ratio, RAR, of at least 1.05, such as 1.08, such as1.1, such as 1.15, such as 1.2, such as 1.25, such as 1.3, such as 1.4,such as 1.5, such as 1.6, such as 1.7, such as 1.8, such as 1.9, such as2, such as 3, such as 4, such as 5 or such as at least 10 compared tothe parent dispersin e.g. a dispersin comprising the polypeptide havingthe amino acid sequence shown in SEQ ID NO: 1, wherein the variant hasat least 60%, at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 95%, such as at least 96%, at least97%, at least 98%, or at least 99%, but less than 100% sequence identityto the polypeptide shown in SEQ ID NO: 1 and preferably wherein thevariant has hexosaminidase e.g. beta-1,6 N-acetylglucosaminidaseactivity.

One preferred aspect of the invention relates to a variant of adispersin parent, wherein the variant comprises one or more motifs GXDE(SEQ ID NO:8), [EQ][NRSHA][YVFL][AGSTC][IVLF][EAQYN][SN] (SEQ IDNO:9),[VIM][LIV]G[GAV]DE[VI][PSA] SEQ ID NO: 10), [WND[SQR][IVL][TLVM](SEQ ID NO: 11), QSTL (SEQ ID NO: 12), NKFFY (SEQ ID NO: 13) or NLD[DR]S(SEQ ID NO: 14), wherein the variant comprises one or more alterations(compared to SEQ ID NO: 1), wherein the alterations are selected fromthe group consisting of: D2A, D2L, D2N, D2R, D2V, D2W, Q3F, Q3I, Q3L,Q3M, Q3P, Q3V, Q3Y, Q3T, S12A, H15F, H15Y, T17W, T17C, T17E, T17F, T17M,T17R, T17V, V18L, E19D, E19K, E19N, E19P, K22A, K22M, K22V, S23A, S23C,S23E, S23I, S23L, S23R, S23T, S23V, L24I, V25R, D26M, Y30*, Y30A, Y30D,Y30L, Y30M, Y30N, Y30R, Y30T, Y30V, G32L, G32M, G32R, N34D, N43*, N43H,N43L, E44*, N45D, N45L, N45V, A49W, A49Y, Y52*, Y52M, G54L, G54M, G54N,S56T, S56W, S57W, E58N, N59A, N59C, N59D, N59E, N59F, N59M, N59R, N59V,N59W, T60V, N62C, N62D, N62H, N62Q, N62W, T63C, T63D, T63L, T63N, T63R,T63V, K67A, K67L, N68L, N68Q, L71H, L71N, L71R, L71V, L71W, S72*, S72C,S72D, S72E, S72F, S72G, S72I, S72M, S72N, S72R, S72T, S72Y, I74L, S77A,D79V, K80*, K80E, K80H, K80L, K80N, K80Q, K80R, K80V, K80W, D81A, D81G,D81L, D81N, D81R, D81S, D81T, D81V, D81W, I82V, L90F, E99Q, E99R, L100S,K103A, K103R, K103V, K104N, K104W, D105N, V106A, V106D, V106E, V106H,V106K, V106L, V106M, V106N, V106Q, V106R, V106W, V106Y, K107A, K107C,K107L, K107M, K107T, K107V, K107W, N110M, N110R, N110V, D111A, D111E,D111M, D111N, D111Q, D111R, D111V, D111W, V113T, T114C, T114S, Y116D,Y116N, Y116R, S117*, S117D, S117H, S117N, S117P, E118*, E118A, E118D,E118G, E118L, E119G, E119W, T120I, T120L, T120M, T120V, T120W, D122*,D122H, D122R, Y123W, Y124C, Y124H, Y124I, Y124K, Y124L, Y124M, Y124N,Y124Q, Y124R, Y124T, Y124V, Y124W, D125C, D125G, D125H, D125K, D125Q,D125R, N126V, R127D, R127H, R127K, R127L, R127M, R127Q, R127W, V128A,V128C, V128D, V128L, V128T, D131V, Q135*, Q135A, Q135D, Q135E, Q135K,Q135M, Q135Y, D138K, D138L, D138M, D138N, D138Q, D138R, D138S, D138V,D138W, E139W, V140I, D142R, D142W, Y145*, Y145H, Y145L, Y145N, Y145V,P147A, P147C, P147D, P147F, P147G, P147L, P147M, P147R, P147S, P147T,P147V, K148A, K148D, K148L, K148V, F149L, F149M, F149N, E150A, E150D,E150H, E150K, E150L, E150M, E150N, E150R, E150V, E150W, E150Y, G151A,G151C, G151D, G151L, G151N, G151P, G151S, G151W, K152D, K152L, K152R,S163P, G164D, G164E, G164H, G164S, G164V, V167A, V167D, V167E, V167L,V167P, V167Q, V167R, V167W, H168N, L170A, L170D, L170E, L170F, L170H,L170K, L170M, L170N, L170P, L170Q, L170R, L170S, L170V, L170W, L170Y,D171A, D171C, D171E, D171K, D171L, D171M, D171Q, D171R, D171V, D171W,D171Y, I173C, D174H, D174M, D174N, D174V, D174W, F175Y, N177M, Q178*,Q178A, Q178K, Q178R, Q178W, I179T, S181C, S181D, S181F, S181G, S181K,S181N, S181P, S181Q, S181T, S181V, S181W, E185A, E185M, E185R, E185V,E185W, S186D, S186E, S186H, S186I, S186K, S186L, S186M, S186N, S186Q,S186R, S186V, S186W, K187C, K187D, K187G, K187R, K187S, K187V, K187W,Y188P, E189L, E189V, E189W, S199C, S199L, S199M, S199Y, E200D, E200F,E200K, E200L, E200M, E200N, E200R, E200W, A203C, A203D, A203E, A203G,A203L, A203M, A203P, A203Q, A203R, A203S, A203T, A203V, A203W, N204L,N204M, N204V, N204W, N204Y, L205I, D207A, D207C, D207E, D207G, D207K,D207N, D207Q, D207R, D207S, D207V, D207W, S208A, S208C, S208D, S208G,S208L, S208Q, S208T, S208V, S208W, S210T, Q215K, Q215R, Q215M, Q215L,Q215*, S217V, T218A, T218L, T218Q, T218R, T218V, S221N, G222D, E224A,E224P, S225G, N227A, N227Q, N227R, N227S, N227T, N227K, D230*, D230N,D230R, D230T, D230W, E232D, E232V, N233D, N233E, N233H, N233Q, N233R,N233W, W234R, G235W, G235A, G235E, G235F, G235H, G235I, G235L, G235M,G235N, G235P, G235S, G235V, S237C, S237G, S237M, S237N, S237W, S237Y,Y244C, Y244E, Y244M, L249H, L249K, L249Q, L249R, L249W, L249Y, S251A,S251L, S251N, S251R, S251W, N252P, N252C, G253D, G253W, F254I, F254L,F254M, F254Y, Q256D, Q256E, Q256R, N260*, N260A, N260C, N260E, N260I,N260K, N260L, N260M, N260Q, N260R, N260T, N260V, N260W, N260Y, E261*,E261A, E261D, E261R, E261W, Q262*, Q262F, Q262H, Q262W, Q262Y, M263K,M263L, M263Q, D264*, D264C, D264E, D264N, Y265F, N267S, N267T, W268C,W268E, W268M, W268R, Y270F, A271D, A271G, H272D, H272I, H272M, H272P,H272V, H272W, N273W, K274R, K274A, K274H, F276A, F276C, F276K, F276N,F276G, F276L, F276M, F276P, F276S, F276V, F276W, I278A, I278K, I278N,I278Q, I278V, S279C, S279D, S279E, S279G, S279N, D280C, D280E, Y281*,Y281A, Y281C, Y281H, Y281K, Y281N, Y281P, Y281R, Y282E, Y282N, H283I,A284I, A284L, A284N, A284P, A284T, A284V, T287N, S288P, S288D, S288K,S288N, V290I, K291L, K291R, K291V, T296C, E300A, E300D, H301C, H301N,H301R, T303A, T303C, T303G, T303K, T303Q, T303R, T303W, D304C, D304M,L305M, L305N, S306C, K308A, K308D, K308E, K308G, K308I, K308L, K308Q,K308S, K308T, K308V, K308Y, K309A, K309C, K309D, K309E, K309G, K309H,K309L, K309M, K309N, K309Q, K309S, K309T, K309I, K312A, K312E, K312L,K312M, K312N, K312Q, K312S, K312W, E314I, E314L, E314V, L315I, L315V,R319A, Y321F and N323R, wherein each position corresponds to theposition of the polypeptide of SEQ ID NO: 1, wherein each substitutionprovides a dispersin variant having an increase in stability measured ashalf-life improvement factor, HIF or residual activity ratio, RAR, of atleast 1.05, such as 1.08, such as 1.1, such as 1.15, such as 1.2, suchas 1.25, such as 1.3, such as 1.4, such as 1.5, such as 1.6, such as1.7, such as 1.8, such as 1.9, such as 2, such as 3, such as 4, such as5 or such as at least 10 compared to the parent dispersin e.g. adispersin comprising the polypeptide having the amino acid sequenceshown in SEQ ID NO: 1, wherein the variant has at least 60%, at least65%, at least 70%, at least 75%, at least 80%, at least 85%, at least90%, at least 95%, such as at least 96%, at least 97%, at least 98%, orat least 99%, but less than 100% sequence identity to the polypeptideshown in SEQ ID NO: 1 and preferably wherein the variant hashexosaminidase e.g. beta-1,6 N-acetylglucosaminidase activity.

One preferred aspect of the invention relates to a variant of adispersin parent, wherein the variant comprises one or more motifs GXDE(SEQ ID NO:8), [EQ][NRSHA][YVFL][AGSTC][IVLF][EAQYN][SN] (SEQ IDNO:9),[VIM][LIV]G[GAV]DE[VI][PSA] SEQ ID NO: 10), [WND[SQR][IVL][TLVM](SEQ ID NO: 11), QSTL (SEQ ID NO: 12), NKFFY (SEQ ID NO: 13) or NLD[DR]S(SEQ ID NO: 14), wherein the variant comprises one or more alterations(compared to SEQ ID NO: 1), wherein the alterations are selected fromthe group consisting of: D2A, D2L, D2N, D2R, D2V, D2W, Q3F, Q3I, Q3L,Q3M, Q3P, Q3V, Q3Y, Q3T, S12A, H15F, H15Y, T17W, T17C, T17E, T17F, T17M,T17R, T17V, V18L, E19D, E19N, E19P, K22A, K22M, K22V, S23C, S23E, S23I,S23L, S23R, S23T, S23V, V25R, D26M, Y30*, Y30D, Y30L, Y30M, Y30N, Y30R,Y30T, Y30V, G32L, G32M, G32R, N43*, N43H, N43L, E44*, N45D, N45L, N45V,A49W, A49Y, Y52*, Y52M, G54L, G54M, G54N, S56T, S56W, S57W, E58N, N59A,N59C, N59D, N59E, N59F, N59M, N59R, N59V, N59W, T60V, N62C, N62D, N62H,N62Q, N62W, T63C, T63D, T63L, T63N, T63R, T63V, K67A, K67L, N68L, N68Q,L71H, L71N, L71R, L71V, L71W, S72*, S72C, S72D, S72E, S72F, S72G, S72I,S72M, S72N, S72R, S72T, S72Y, I74L, S77A, D79V, K80*, K80E, K80H, K80L,K80N, K80Q, K80V, K80W, D81A, D81G, D81L, D81R, D81S, D81T, D81V, D81W,I82V, L90F, E99Q, E99R, L100S, K103A, K103R, K104N, K104W, D105N, V106A,V106D, V106E, V106H, V106K, V106L, V106M, V106N, V106Q, V106W, V106Y,K107A, K107C, K107L, K107M, K107T, K107V, K107W, N110M, N110R, N110V,D111A, D111E, D111M, D111N, D111Q, D111R, D111V, D111W, V113T, T114C,T114S, Y116D, Y116N, Y116R, S117*, S117D, S117H, S117N, S117P, E118*,E118A, E118D, E118G, E118L, E119G, E119W, T120I, T120L, T120M, T120V,T120W, D122*, D122H, D122R, Y123W, Y124C, Y124I, Y124K, Y124L, Y124M,Y124Q, Y124R, Y124T, Y124V, Y124W, D125C, D125G, D125K, D125Q, D125R,N126V, R127D, R127H, R127K, R127L, R127M, R127Q, R127W, V128C, V128L,V128T, D131V, Q135*, Q135A, Q135D, Q135E, Q135K, Q135M, Q135Y, D138K,D138L, D138M, D138Q, D138R, D138S, D138V, D138W, E139W, D142R, D142W,Y145*, Y145H, Y145L, Y145N, Y145V, P147A, P147C, P147D, P147F, P147G,P147L, P147M, P147R, P147S, P147T, P147V, K148A, K148D, K148L, K148V,F149L, F149M, F149N, E150D, E150H, E150K, E150L, E150M, E150N, E150R,E150V, E150W, E150Y, G151A, G151C, G151D, G151L, G151N, G151P, G151S,G151W, K152D, K152L, K152R, G164D, G164E, G164H, G164S, G164V, V167D,V167E, V167L, V167P, V167Q, V167R, V167W, H168N, L170A, L170D, L170E,L170F, L170H, L170K, L170M, L170N, L170P, L170Q, L170R, L170S, L170V,L170W, L170Y, D171A, D171C, D171E, D171K, D171L, D171M, D171Q, D171R,D171V, D171W, D171Y, I173C, D174H, D174M, D174N, D174V, D174W, F175Y,N177M, Q178*, Q178A, Q178K, Q178R, Q178W, I179T, S181C, S181D, S181F,S181G, S181N, S181P, S181Q, S181T, S181V, S181W, E185M, E185R, E185V,E185W, S186D, S186E, S186H, S186I, S186K, S186L, S186M, S186N, S186Q,S186R, S186V, S186W, K187C, K187D, K187G, K187R, K187S, K187V, K187W,Y188P, E189L, E189V, E189W, S199C, S199L, S199M, S199Y, E200D, E200F,E200K, E200L, E200M, E200N, E200R, E200W, A203C, A203D, A203E, A203G,A203L, A203M, A203P, A203R, A203S, A203T, A203V, A203W, N204L, N204M,N204V, N204W, N204Y, L205I, D207A, D207C, D207E, D207G, D207N, D207Q,D207S, D207V, D207W, S208A, S208C, S208D, S208G, S208L, S208Q, S208T,S208V, S208W, S210T, Q215R, Q215M, Q215L, Q215*, S217V, T218A, T218L,T218Q, T218R, T218V, G222D, E224A, E224P, N227A, N227Q, N227R, N227S,N227T, N227K, D230*, D230R, D230T, D230W, E232D, E232V, N233H, N233Q,N233R, N233W, W234R, G235W, G235A, G235E, G235F, G235H, G235I, G235L,G235M, G235N, G235P, G235S, G235V, S237C, S237G, S237M, S237N, S237W,S237Y, Y244C, Y244E, Y244M, L249H, L249K, L249Q, L249R, L249W, L249Y,S251L, S251N, S251R, S251W, N252P, N252C, G253D, G253W, F254I, F254L,F254M, F254Y, Q256E, Q256R, N260*, N260A, N260C, N260E, N260I, N260K,N260L, N260M, N260Q, N260R, N260V, N260W, N260Y, E261*, E261A, E261D,E261R, E261W, Q262*, Q262F, Q262H, Q262W, Q262Y, M263K, M263L, M263Q,D264*, D264C, D264E, Y265F, N267S, N267T, W268C, W268E, W268M, W268R,Y270F, A271D, A271G, H272D, H272I, H272M, H272P, H272V, H272W, N273W,K274R, K274A, K274H, F276A, F276C, F276K, F276N, F276G, F276L, F276M,F276P, F276S, F276V, F276W, I278A, I278K, I278N, I278Q, I278V, S279C,S279D, S279E, S279G, S279N, D280C, D280E, Y281*, Y281A, Y281C, Y281H,Y281K, Y281N, Y281P, Y281R, Y282E, Y282N, H283I, A284I, A284L, A284N,A284P, A284T, A284V, T287N, S288D, S288K, S288N, V290I, K291L, K291R,K291V, T296C, E300A, E300D, H301C, H301N, H301R, T303A, T303C, T303G,T303K, T303Q, T303R, T303W, D304C, D304M, L305M, L305N, S306C, K308A,K308D, K308G, K308I, K308L, K308Q, K308S, K308T, K308V, K308Y, K309A,K309C, K309D, K309H, K309L, K309M, K309N, K309Q, K309S, K309T, K309I,K312A, K312L, K312M, K312N, K312Q, K312S, K312W, E314I, E314L, E314V,L315I, L315V, R319A, and N323R, wherein each position corresponds to theposition of the polypeptide of SEQ ID NO: 1, wherein each substitutionprovides a dispersin variant having an increase in stability measured ashalf-life improvement factor, HIF or residual activity ratio, RAR, of atleast 1.05, such as 1.08, such as 1.1, such as 1.15, such as 1.2, suchas 1.25, such as 1.3, such as 1.4, such as 1.5, such as 1.6, such as1.7, such as 1.8, such as 1.9, such as 2, such as 3, such as 4, such as5 or such as at least 10 compared to the parent dispersin e.g. adispersin comprising the polypeptide having the amino acid sequenceshown in SEQ ID NO: 1, wherein the variant has at least 60%, at least65%, at least 70%, at least 75%, at least 80%, at least 85%, at least90%, at least 95%, such as at least 96%, at least 97%, at least 98%, orat least 99%, but less than 100% sequence identity to the polypeptideshown in SEQ ID NO: 1 and preferably wherein the variant hashexosaminidase e.g. beta-1,6 N-acetylglucosaminidase activity.

One preferred aspect of the invention relates to a variant of adispersin parent, wherein the variant comprises one or more motifs GXDE(SEQ ID NO:8), [EQ][NRSHA][YVFL][AGSTC][IVLF][EAQYN][SN] (SEQ IDNO:9),[VIM][LIV]G[GAV]DE[VI][PSA] SEQ ID NO: 10), [WND[SQR][IVL][TLVM](SEQ ID NO: 11), QSTL (SEQ ID NO: 12), NKFFY (SEQ ID NO: 13) or NLD[DR]S(SEQ ID NO: 14), wherein the variant comprises one or more alterations(compared to SEQ ID NO: 1), wherein the alterations are selected fromthe group consisting of: D2V, Q3F, Q3I, S12A, H15Y, T17W, T17E, V18L,K22M, S23I, S23C, S23T, S23L, S23V, S23E, V25R, Y30L, A49W, A49Y, S56T,N59D, N59C, N59E, N59R, N59F, N59W, N59V, N62C, N62D, T63C, N68Q, S72D,S72E, I74L, S77A, I82V, L90F, E99Q, L100S, T114S, T114C, Y123W, Y124I,Y124M, Y124R, Y124V, Y124Q, Y124T, Y124K, D125R, D125C, D125G, D125K,D125Q, Q135M, D138R, D138K, D138Q, L170D, L170K, L170S, L170H, D171E,D171K, D171Y, D171M, D171Q, D171L, I173C, D174W, D174H, D174M, D174N,F175Y, Q178K, I179T, S181T, S181F, S181Q, S181G, S181N, S181C, E185R,E185M, E185V, S186K, S186M, S186R, S186H, K187G, Y188P, E189V, S199C,S199L, A203G, A203E, A203V, N204L, N204Y, N204V, L205I, D207N, D207S,D207C, D207G, S210T, Q215R, T218Q, N227T, N227K, E232D, G235W, S237W,Y244M, Y244C, N252C, Q256E, N260Q, Q262H, M263Q, D264E, Y265F, N267T,N267S, Y270F, H272M, H272P, H272I, H272V, N273W, K274H, F276A, F276N,F276K, F276C, I278V, S279N, S279D, S279G, D280E, D280C, Y281P, Y282N,H283I, A284T, A284L, A284I, S288K, V290I, K291R, T296C, T303Q, D304M,D304C, L305M, L305N, S306C, K308D, K308A, K308V, K308Q, K308S, K308Y,K308G, K308L, K308T, K308I, K309C, K309L, K309D, K309Q, K309N, K309T,K309A, K309S, K309M, K309H, K312A, K312Q, K312S, K312W, K312L, K312N,E314L, E314V, E314I, L315I, R319A and N323R, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1, whereineach substitution provides a dispersin variant having an increase instability measured as half-life improvement factor, HIF or residualactivity ratio, RAR, of at least 1.05, such as 1.08, such as 1.1, suchas 1.15, such as 1.2, such as 1.25, such as 1.3, such as 1.4, such as1.5, such as 1.6, such as 1.7, such as 1.8, such as 1.9, such as 2, suchas 3, such as 4, such as 5 or such as at least 10 compared to the parentdispersin e.g. a dispersin comprising the polypeptide having the aminoacid sequence shown in SEQ ID NO: 1, wherein the variant has at least60%, at least 65%, at least 70%, at least 75%, at least 80%, at least85%, at least 90%, at least 95%, such as at least 96%, at least 97%, atleast 98%, or at least 99%, but less than 100% sequence identity to thepolypeptide shown in SEQ ID NO: 1 and preferably wherein the variant hashexosaminidase e.g. beta-1,6 N-acetylglucosaminidase activity.

Addition of any one of the mutations of the invention, have shown toimprove stability.

In some preferred aspects, a dispersin variant of the inventioncomprises 2, 3, 4, 5, 6, 7, 8 or 9 or more substitutions (compared toSEQ ID NO: 1), wherein the substitutions are selected from the groupconsisting of: D2V, Q3F, Q3I, S12A, H15Y, T17W, T17E, V18L, K22M, S23I,S23C, S23T, S23L, S23V, S23E, V25R, Y30L, A49W, A49Y, S56T, N59D, N59C,N59E, N59R, N59F, N59W, N59V, N62C, N62D, T63C, N68Q, S72D, S72E, I74L,S77A, I82V, L90F, E99Q, L100S, T114S, T114C, Y123W, Y124I, Y124M, Y124R,Y124V, Y124Q, Y124T, Y124K, D125R, D125C, D125G, D125K, D125Q, Q135M,D138R, D138K, D138Q, L170D, L170K, L170S, L170H, D171E, D171K, D171Y,D171M, D171Q, D171L, I173C, D174W, D174H, D174M, D174N, F175Y, Q178K,I179T, S181T, S181F, S181Q, S181G, S181N, S181C, E185R, E185M, E185V,S186K, S186M, S186R, S186H, K187G, Y188P, E189V, S199C, S199L, A203G,A203E, A203V, N204L, N204Y, N204V, L205I, D207N, D207S, D207C, D207G,S210T, Q215R, T218Q, S225G, N227T, N227K, E232D, G235W, S237W, Y244M,Y244C, N252C, Q256E, N260Q, Q262H, M263Q, D264E, Y265F, N267T, N267S,Y270F, H272M, H272P, H272I, H272V, N273W, K274H, F276A, F276N, F276K,F276C, I278V, S279N, S279D, S279G, D280E, D280C, Y281P, Y282N, H283I,A284T, A284L, A284I, S288K, V290I, K291R, T296C, T303Q, D304M, D304C,L305M, L305N, S306C, K308D, K308A, K308V, K308Q, K308S, K308Y, K308G,K308L, K308T, K308I, K309C, K309L, K309D, K309Q, K309N, K309T, K309A,K309S, K309M, K309H, K312A, K312Q, K312S, K312W, K312L, K312N, E314L,E314V, E314I, L315I, R319A and N323R, wherein the variant has at least60%, at least 65%, at least 70%, at least 75%, at least 80%, at least85%, at least 90%, at least 95%, such as at least 96%, at least 97%, atleast 98%, or at least 99%, but less than 100%, sequence identity to thepolypeptide shown in SEQ ID NO: 1, wherein each position corresponds tothe position of the polypeptide shown in SEQ ID NO: 1 (numberingaccording to SEQ ID NO: 1) and wherein the variant has at least oneimproved property compared to the parent dispersin e.g. a dispersincomprising the polypeptide having the amino acid sequence shown in SEQID NO: 1, preferably the improved property is, improved stability,wherein stability is tested as described in Example 4 and Example 5,preferably wherein the variant has hexosaminidase e.g. beta-1,6N-acetylglucosaminidase activity.

In some aspects, the dispersin variant of the invention has an improvedstability, measured as Half-life Improvement Factor, HIF, compared tothe parent or compared to the dispersin having the polypeptide shown inSEQ ID NO: 1 or compared to a dispersin having the identical amino acidsequence of the dispersin variant but not having the alterations at 2,3, 4, 5, 6, 7, 8 or 9 or more of the specified positions.

In some aspects, the dispersin variant of the invention has an improvedstability, measured as residual activity ratio, RAR, compared to theparent or compared to the dispersin having the polypeptide shown in SEQID NO: 1 or compared to a dispersin having the identical amino acidsequence of the dispersin variant but not having the alterations at 2,3, 4, 5, 6, 7, 8 or 9 or more of the specified positions.

In one preferred embodiment of the invention the dispersin variant ofthe invention comprises one or more of the following substitution(s),Q3F, Q3I, H15Y, T17W , A49W, N59E, V140I, S163P, S186R, D207N, Q215K,T218Q, S225G, N227T, E232D, G235W, S237W, N252P, N260Q, N267T, H272V,H272P, S279D, Y281P, S288P, K308Q, K309E or K312Q, wherein the varianthas at least 60%, at least 65%, at least 70%, at least 75%, at least80%, at least 85%, at least 90%, at least 95%, such as at least 96%, atleast 97%, at least 98%, or at least 99%, but less than 100% sequenceidentity to the polypeptide shown in SEQ ID NO: 1, wherein each positioncorresponds to the position of the polypeptide shown in SEQ ID NO: 1(numbering according to SEQ ID NO: 1).

In one preferred embodiment of the invention the dispersin variant ofthe invention comprises one or more of the following substitution(s),Q3I, H15Y, A49W, N59E, S163P, S186R, S225G, N227T, E232D, G235W, N252P,N260Q, H272V, S279D, Y281P, K308Q, K309E or K312Q, wherein the varianthas at least 60%, at least 65%, at least 70%, at least 75%, at least80%, at least 85%, at least 90%, at least 95%, such as at least 96%, atleast 97%, at least 98%, or at least 99%, but less than 100% sequenceidentity to the polypeptide shown in SEQ ID NO: 1, wherein each positioncorresponds to the position of the polypeptide shown in SEQ ID NO: 1(numbering according to SEQ ID NO: 1).

In some preferred embodiment of the invention the dispersin variant ofthe invention comprises one or more of the following substitution sets:Q3I+A49W, Q3I+N59E, Q3I+S163P, Q3I+S186R, Q3I+Q215K, Q3I+S225G,Q3I+N227T, Q3I+N252P, Q3I+N267T, Q3I+F276A, Q3I+Y281P, Q3I+K308Q,Q3I+K308E, Q3I+K309E, Q3I+K312Q, Q3I+K312E, Q3F+A49W, Q3F+N59E,Q3F+S163P, Q3F+S186R, Q3F+Q215K, Q3F+S225G, Q3F+N227T, Q3F+N252P,Q3F+N267T, Q3F+F276A, Q3F+Y281P, Q3F+K308Q, Q3F+K308E, Q3F+K309E,Q3F+K312E, Q3F+K312Q, A49W+N59E, A49W+S163P, A49W+S186R, A49W+Q215K,A49W+S225G, A49W+N227T, A49W+N252P, A49W+N267T, A49W+F276A, A49W+Y281P,A49W+K308Q, A49W+K308E, A49W+K309E, A49W+K312Q, A49W+K312E, N59E+S163P,N59E+S186R, N59E+Q215K, N59E+S225G, N59E+N227T, N59E+N252P, N59E+N267T,N59E+F276A, N59E+Y281P, N59E+K308Q, N59E+K308E, N59E+K309E, N59E+K312Q,N59E+K312E, S163P+S186R, S163P+Q215K, S163P+S225G, S163P+N227T,S163P+N252P, S163P+N267T, S163P+F276A, S163P+Y281P, S163P+K308E,S163P+K308Q, S163P+K309E, S163P+K312Q, S163P+K312E, S186R+Q215K,S186R+S225G, S186R+N227T, S186R+N252P, S186R+N267T, S186R+F276A,S186R+Y281P, S186R+K308E, S186R+K308Q, S186R+K309E, S186R+K312Q,S186R+K312E, Q215K+S225G, Q215K+N227T, Q215K+N252P, Q215K+N267T,Q215K+F276A, Q215K+Y281P, Q215K+K308E, Q215K+K308Q, Q215K+K309E,Q215K+K312Q, Q215K+K312E, S225G+N227T, S225G+N252P, S225G+N267T,S225G+F276A, S225G+Y281P, S225G+K308E, S225G+K308Q, S225G+K309E,S225G+K312Q, S225G+K312E, N227T+N252P, N227T+N267T, N227T+F276A,N227T+Y281P, N227T+K308Q, N227T+K308E, N227T+K309E, N227T+K312QN227T+K312E, N252P+N267T, N252P+F276A, N252P+Y281P, N252P+K308Q,N252P+K308E, N252P+K309E, N252P+K312Q, N252P+K312E, N267T+F276A,N267T+Y281P, N267T+K308E, N267T+K309E, N267T+K312Q, N267T+K312E,F276A+Y281P, F276A+K308Q, F276A+K308E, F276A+K309E, F276A+K312Q,F276A+K312E, Y281P+K308Q, Y281P+K308E, Y281P+K309E, Y281P+K312Q,Y281P+K312E, K308E+K312Q, K308E+K312E, K308Q+K312Q, K309E+K312Q orK309E+K312E, wherein the variant has at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100% sequence identity to the polypeptide shown inSEQ ID NO: 1, wherein each position corresponds to the position of thepolypeptide shown in SEQ ID NO: 1 (numbering according to SEQ ID NO: 1).

In one preferred embodiment the dispersin variant of the inventioncomprises at least two substitutions wherein the substitutions orsubstitution sets are selected from the group consisting of:

F276A+K308E, F276A+K309E, K308E+K309E, K308E+K312E, K309E+K312E,N227T+F276A, N227T+K308E, N227T+K312E, N227T+N252P, N227T+N267T,N252P+F276A, N252P+K308E, N252P+K309E, N252P+N267T, N267T+F276A,N267T+K308E, N267T+K309E, N267T+K312E, Q215K+F276A, Q215K+K308E,Q215K+K309E, Q215K+K312E, Q215K+N227T, Q215K+N252P, Q215K+N267T,S163P+F276A, S163P+K308E, S163P+K309E, S163P+K312E, S163P+N252P,S163P+N267T and S163P+Q215K.

In one preferred embodiment the dispersin variant of the inventioncomprises at least two substitutions, wherein the substitution sets areselected from the group consisting of: Q3I+H15Y, Q3I+A49W, Q3I+N59E,Q3I+S163P, Q3I+S186R, Q3I+S225G, Q3I+N227T, Q3I+E232D, Q3I+G235W,Q3I+N252P, Q3I+N260Q, Q3I+H272V, Q3I+S279D, Q3I+Y281P, Q3I+K308Q,Q3I+K309E, Q3I+K312Q, H15Y+A49W, H15Y+N59E, H15Y+S163P, H15Y+S186R,H15Y+S225G, H15Y+N227T, H15Y+E232D, H15Y+G235W, H15Y+N252P, H15Y+N260Q,H15Y+H272V, H15Y+S279D, H15Y+Y281P, H15Y+K308Q, H15Y+K309E, H15Y+K312Q,A49W+N59E, A49W+S163P, A49W+S186R, A49W+S225G, A49W+N227T, A49W+E232D,A49W+G235W, A49W+N252P, A49W+N260Q, A49W+H272V, A49W+S279D, A49W+Y281P,A49W+K308Q, A49W+K309E, A49W+K312Q, N59E+S163P, N59E+S186R, N59E+S225G,N59E+N227T, N59E+E232D, N59E+G235W, N59E+N252P, N59E+N260Q, N59E+H272V,N59E+S279D, N59E+Y281P, N59E+K308Q, N59E+K309E, N59E+K312Q, S163P+S186R,S163P+S225G, S163P+N227T, S163P+E232D, S163P+G235W, S163P+N252P,S163P+N260Q, S163P+H272V, S163P+S279D, S163P+Y281P, S163P+K308Q,S163P+K309E, S163P+K312Q, S186R+S225G, S186R+N227T, S186R+E232D,S186R+G235W, S186R+N252P, S186R+N260Q, S186R+H272V, S186R+S279D,S186R+Y281P, S186R+K308Q, S186R+K309E, S186R+K312Q, S225G+N227T,S225G+E232D, S225G+G235W, S225G+N252P, S225G+N260Q, S225G+H272V,S225G+S279D, S225G+Y281P, S225G+K308Q, S225G+K309E, S225G+K312Q,N227T+E232D, N227T+G235W, N227T+N252P, N227T+N260Q, N227T+H272V,N227T+S279D, N227T+Y281P, N227T+K308Q, N227T+K309E, N227T+K312Q,E232D+G235W, E232D+N252P, E232D+N260Q, E232D+H272V, E232D+S279D,E232D+Y281P, E232D+K308Q, E232D+K309E, E232D+K312Q, G235W+N252P,G235W+N260Q, G235W+H272V, G235W+S279D, G235W+Y281P, G235W+K308Q,G235W+K309E, G235W+K312Q, N252P+N260Q, N252P+H272V, N252P+S279D,N252P+Y281P, N252P+K308Q, N252P+K309E, N252P+K312Q, N260Q+H272V,N260Q+S279D, N260Q+Y281P, N260Q+K308Q, N260Q+K309E, N260Q+K312Q,H272V+S279D, H272V+Y281P, H272V+K308Q, H272V+K309E, H272V+K312Q,S279D+Y281P, S279D+K308Q, S279D+K309E, S279D+K312Q, Y281P+K308Q,Y281P+K309E, Y281P+K312Q, K308Q+K309E, K308Q+K312Q and K309E+K312Q.

In one preferred embodiment the dispersin variant of the inventioncomprises at least three substitutions, wherein the substitutions orsubstitution sets are selected from the group consisting of:

K312E+N227T+F276A, K312E+N227T+K308E, K312E+N227T+K309E,K312E+N227T+K312E, K312E+F276A+K308E, K312E+F276A+K309E,K312E+F276A+K312E, K312E+K308E+K309E, K312E+K308E+K312E,K312E+K309E+K312E, N227T+F276A+K308E, N227T+F276A+K309E,N227T+F276A+K312E, N227T+K308E+K309E, N227T+K308E+K312E,N227T+K309E+K312E, F276A+K308E+K309E, F276A+K308E+K312E,F276A+K309E+K312E, and K308E+K309E+K312E.

In one preferred embodiment the dispersin variant of the inventioncomprises at least three substitutions, wherein the substitution setsare selected from the group consisting of:

Q3I+H15Y+A49W, Q3I+H15Y+N59E, Q3I+H15Y+S163P, Q3I+H15Y+S186R,Q3I+H15Y+S225G, Q3I+H15Y+N227T, Q3I+H15Y+E232D, Q3I+H15Y+G235W,Q3I+H15Y+N252P, Q3I+H15Y+N260Q, Q3I+H15Y+H272V, Q3I+H15Y+S279D,Q3I+H15Y+Y281P, Q3I+H15Y+K308Q, Q3I+H15Y+K309E, Q3I+H15Y+K312Q,Q3I+A49W+N59E, Q3I+A49W+S163P, Q3I+A49W+S186R, Q3I+A49W+S225G,Q3I+A49W+N227T, Q3I+A49W+E232D, Q3I+A49W+G235W, Q3I+A49W+N252P,Q3I+A49W+N260Q, Q3I+A49W+H272V, Q3I+A49W+S279D, Q3I+A49W+Y281P,Q3I+A49W+K308Q, Q3I+A49W+K309E, Q3I+A49W+K312Q, Q3I+N59E+S163P,Q3I+N59E+S186R, Q3I+N59E+S225G, Q3I+N59E+N227T, Q3I+N59E+E232D,Q3I+N59E+G235W, Q3I+N59E+N252P, Q3I+N59E+N260Q, Q3I+N59E+H272V,Q3I+N59E+S279D, Q3I+N59E+Y281P, Q3I+N59E+K308Q, Q3I+N59E+K309E,Q3I+N59E+K312Q, Q3I+S163P+S186R, Q3I+S163P+S225G, Q3I+S163P+N227T,Q3I+S163P+E232D, Q3I+S163P+G235W, Q3I+S163P+N252P, Q3I+S163P+N260Q,Q3I+S163P+H272V, Q3I+S163P+S279D, Q3I+S163P+Y281P, Q3I+S163P+K308Q,Q3I+S163P+K309E, Q3I+S163P+K312Q, Q3I+S186R+S225G, Q3I+S186R+N227T,Q3I+S186R+E232D, Q3I+S186R+G235W, Q3I+S186R+N252P, Q3I+S186R+N260Q,Q3I+S186R+H272V, Q3I+S186R+S279D, Q3I+S186R+Y281P, Q3I+S186R+K308Q,Q3I+S186R+K309E, Q3I+S186R+K312Q, Q3I+S225G+N227T, Q3I+S225G+E232D,Q3I+S225G+G235W, Q3I+S225G+N252P, Q3I+S225G+N260Q, Q3I+S225G+H272V,Q3I+S225G+S279D, Q3I+S225G+Y281P, Q3I+S225G+K308Q, Q3I+S225G+K309E,Q3I+S225G+K312Q, Q3I+N227T+E232D, Q3I+N227T+G235W, Q3I+N227T+N252P,Q3I+N227T+N260Q, Q3I+N227T+H272V, Q3I+N227T+S279D, Q3I+N227T+Y281P,Q3I+N227T+K308Q, Q3I+N227T+K309E, Q3I+N227T+K312Q, Q3I+E232D+G235W,Q3I+E232D+N252P, Q3I+E232D+N260Q, Q3I+E232D+H272V, Q3I+E232D+S279D,Q3I+E232D+Y281P, Q3I+E232D+K308Q, Q3I+E232D+K309E, Q3I+E232D+K312Q,Q3I+G235W+N252P, Q3I+G235W+N260Q, Q3I+G235W+H272V, Q3I+G235W+S279D,Q3I+G235W+Y281P, Q3I+G235W+K308Q, Q3I+G235W+K309E, Q3I+G235W+K312Q,Q3I+N252P+N260Q, Q3I+N252P+H272V, Q3I+N252P+S279D, Q3I+N252P+Y281P,Q3I+N252P+K308Q, Q3I+N252P+K309E, Q3I+N252P+K312Q, Q3I+N260Q+H272V,Q3I+N260Q+S279D, Q3I+N260Q+Y281P, Q3I+N260Q+K308Q, Q3I+N260Q+K309E,Q3I+N260Q+K312Q, Q3I+H272V+S279D, Q3I+H272V+Y281P, Q3I+H272V+K308Q,Q3I+H272V+K309E, Q3I+H272V+K312Q, Q3I+S279D+Y281P, Q3I+S279D+K308Q,Q3I+S279D+K309E, Q3I+S279D+K312Q, Q3I+Y281P+K308Q, Q3I+Y281P+K309E,Q3I+Y281P+K312Q, Q3I+K308Q+K309E, Q3I+K308Q+K312Q, Q3I+K309E+K312Q,H15Y+A49W+N59E, H15Y+A49W+S163P, H15Y+A49W+S186R, H15Y+A49W+S225G,H15Y+A49W+N227T, H15Y+A49W+E232D, H15Y+A49W+G235W, H15Y+A49W+N252P,H15Y+A49W+N260Q, H15Y+A49W+H272V, H15Y+A49W+S279D, H15Y+A49W+Y281P,H15Y+A49W+K308Q, H15Y+A49W+K309E, H15Y+A49W+K312Q, H15Y+N59E+S163P,H15Y+N59E+S186R, H15Y+N59E+S225G, H15Y+N59E+N227T, H15Y+N59E+E232D,H15Y+N59E+G235W, H15Y+N59E+N252P, H15Y+N59E+N260Q, H15Y+N59E+H272V,H15Y+N59E+S279D, H15Y+N59E+Y281P, H15Y+N59E+K308Q, H15Y+N59E+K309E,H15Y+N59E+K312Q, H15Y+S163P+S186R, H15Y+S163P+S225G, H15Y+S163P+N227T,H15Y+S163P+E232D, H15Y+S163P+G235W, H15Y+S163P+N252P, H15Y+S163P+N260Q,H15Y+S163P+H272V, H15Y+S163P+S279D, H15Y+S163P+Y281P, H15Y+S163P+K308Q,H15Y+S163P+K309E, H15Y+S163P+K312Q, H15Y+S186R+S225G, H15Y+S186R+N227T,H15Y+S186R+E232D, H15Y+S186R+G235W, H15Y+S186R+N252P, H15Y+S186R+N260Q,H15Y+S186R+H272V, H15Y+S186R+S279D, H15Y+S186R+Y281P, H15Y+S186R+K308Q,H15Y+S186R+K309E, H15Y+S186R+K312Q, H15Y+S225G+N227T, H15Y+S225G+E232D,H15Y+S225G+G235W, H15Y+S225G+N252P, H15Y+S225G+N260Q, H15Y+S225G+H272V,H15Y+S225G+S279D, H15Y+S225G+Y281P, H15Y+S225G+K308Q, H15Y+S225G+K309E,H15Y+S225G+K312Q, H15Y+N227T+E232 D, H15Y+N227T+G235W, H15Y+N227T+N252P,H15Y+N227T+N260Q, H15Y+N227T+H272V, H15Y+N227T+S279D, H15Y+N227T+Y281P,H15Y+N227T+K308Q, H15Y+N227T+K309E, H15Y+N227T+K312Q, H15Y+E232 D+G235W,H15Y+E232 D+N252P, H15Y+E232D+N260Q, H15Y+E232D+H272V, H15Y+E232D+S279D, H15Y+E232 D+Y281P, H15Y+E232D+K308Q, H15Y+E232 D+K309E,H15Y+E232D+K312Q, H15Y+G235W+N252P, H15Y+G235W+N260Q, H15Y+G235W+H272V,H15Y+G235W+S279D, H15Y+G235W+Y281P, H15Y+G235W+K308Q, H15Y+G235W+K309E,H15Y+G235W+K312Q, H15Y+N252P+N260Q, H15Y+N252P+H272V, H15Y+N252P+S279D,H15Y+N252P+Y281P, H15Y+N252P+K308Q, H15Y+N252P+K309E, H15Y+N252P+K312Q,H15Y+N260Q+H272V, H15Y+N260Q+S279D, H15Y+N260Q+Y281P, H15Y+N260Q+K308Q,H15Y+N260Q+K309E, H15Y+N260Q+K312Q, H15Y+H272V+S279D, H15Y+H272V+Y281P,H15Y+H272V+K308Q, H15Y+H272V+K309E, H15Y+H272V+K312Q, H15Y+S279D+Y281P,H15Y+S279D+K308Q, H15Y+S279D+K309E, H15Y+S279D+K312Q, H15Y+Y281P+K308Q,H15Y+Y281P+K309E, H15Y+Y281P+K312Q, H15Y+K308Q+K309E, H15Y+K308Q+K312Q,H15Y+K309E+K312Q, A49W+N59E+S163P, A49W+N59E+S186R, A49W+N59E+S225G,A49W+N59E+N227T, A49W+N59E+E232D, A49W+N59E+G235W, A49W+N59E+N252P,A49W+N59E+N260Q, A49W+N59E+H272V, A49W+N59E+S279D, A49W+N59E+Y281P,A49W+N59E+K308Q, A49W+N59E+K309E, A49W+N59E+K312Q, A49W+S163P+S186R,A49W+S163P+S225G, A49W+S163P+N227T, A49W+S163P+E232D, A49W+S163P+G235W,A49W+S163P+N252P, A49W+S163P+N260Q, A49W+S163P+H272V, A49W+S163P+S279D,A49W+S163P+Y281P, A49W+S163P+K308Q, A49W+S163P+K309E, A49W+S163P+K312Q,A49W+S186R+S225G, A49W+S186R+N227T, A49W+S186R+E232D, A49W+S186R+G235W,A49W+S186R+N252P, A49W+S186R+N260Q, A49W+S186R+H272V, A49W+S186R+S279D,A49W+S186R+Y281P, A49W+S186R+K308Q, A49W+S186R+K309E, A49W+S186R+K312Q,A49W+S225G+N227T, A49W+S225G+E232D, A49W+S225G+G235W, A49W+S225G+N252P,A49W+S225G+N260Q, A49W+S225G+H272V, A49W+S225G+S279D, A49W+S225G+Y281P,A49W+S225G+K308Q, A49W+S225G+K309E, A49W+S225G+K312Q, A49W+N227T+E232D,A49W+N227T+G235W, A49W+N227T+N252P, A49W+N227T+N260Q, A49W+N227T+H272V,A49W+N227T+S279D, A49W+N227T+Y281P, A49W+N227T+K308Q, A49W+N227T+K309E,A49W+N227T+K312Q, A49W+E232D+G235W, A49W+E232D+N252P, A49W+E232D+N260Q,A49W+E232D+H272V, A49W+E232D+S279D, A49W+E232D+Y281P, A49W+E232D+K308Q,A49W+E232 D+K309E, A49W+E232D+K312Q, A49W+G235W+N252P, A49W+G235W+N260Q,A49W+G235W+H272V, A49W+G235W+S279D, A49W+G235W+Y281P, A49W+G235W+K308Q,A49W+G235W+K309E, A49W+G235W+K312Q, A49W+N252P+N260Q, A49W+N252P+H272V,A49W+N252P+S279D, A49W+N252P+Y281P, A49W+N252P+K308Q, A49W+N252P+K309E,A49W+N252P+K312Q, A49W+N260Q+H272V, A49W+N260Q+S279D, A49W+N260Q+Y281P,A49W+N260Q+K308Q, A49W+N260Q+K309E, A49W+N260Q+K312Q, A49W+H272V+S279D,A49W+H272V+Y281P, A49W+H272V+K308Q, A49W+H272V+K309E, A49W+H272V+K312Q,A49W+S279D+Y281P, A49W+S279D+K308Q, A49W+S279D+K309E, A49W+S279D+K312Q,A49W+Y281P+K308Q, A49W+Y281P+K309E, A49W+Y281P+K312Q, A49W+K308Q+K309E,A49W+K308Q+K312Q, A49W+K309E+K312Q, N59E+S163P+S186R, N59E+S163P+S225G,N59E+S163P+N227T, N59E+S163P+E232D, N59E+S163P+G235W, N59E+S163P+N252P,N59E+S163P+N260Q, N59E+S163P+H272V, N59E+S163P+S279D, N59E+S163P+Y281P,N59E+S163P+K308Q, N59E+S163P+K309E, N59E+S163P+K312Q, N59E+S186R+S225G,N59E+S186R+N227T, N59E+S186R+E232D, N59E+S186R+G235W, N59E+S186R+N252P,N59E+S186R+N260Q, N59E+S186R+H272V, N59E+S186R+S279D, N59E+S186R+Y281P,N59E+S186R+K308Q, N59E+S186R+K309E, N59E+S186R+K312Q, N59E+S225G+N227T,N59E+S225G+E232D, N59E+S225G+G235W, N59E+S225G+N252P, N59E+S225G+N260Q,N59E+S225G+H272V, N59E+S225G+S279D, N59E+S225G+Y281P, N59E+S225G+K308Q,N59E+S225G+K309E, N59E+S225G+K312Q, N59E+N227T+E232 D, N59E+N227T+G235W,N59E+N227T+N252P, N59E+N227T+N260Q, N59E+N227T+H272V, N59E+N227T+S279D,N59E+N227T+Y281P, N59E+N227T+K308Q, N59E+N227T+K309E, N59E+N227T+K312Q,N59E+E232 D+G235W, N59E+E232 D+N252P, N59E+E232D+N260Q,N59E+E232D+H272V, N59E+E232 D+S279D, N59E+E232 D+Y281P,N59E+E232D+K308Q, N59E+E232 D+K309E, N59E+E232D+K312Q, N59E+G235W+N252P,N59E+G235W+N260Q, N59E+G235W+H272V, N59E+G235W+S279D, N59E+G235W+Y281P,N59E+G235W+K308Q, N59E+G235W+K309E, N59E+G235W+K312Q, N59E+N252P+N260Q,N59E+N252P+H272V, N59E+N252P+S279D, N59E+N252P+Y281P, N59E+N252P+K308Q,N59E+N252P+K309E, N59E+N252P+K312Q, N59E+N260Q+H272V, N59E+N260Q+S279D,N59E+N260Q+Y281P, N59E+N260Q+K308Q, N59E+N260Q+K309E, N59E+N260Q+K312Q,N59E+H272V+S279D, N59E+H272V+Y281P, N59E+H272V+K308Q, N59E+H272V+K309E,N59E+H272V+K312Q, N59E+S279D+Y281P, N59E+S279D+K308Q, N59E+S279D+K309E,N59E+S279D+K312Q, N59E+Y281P+K308Q, N59E+Y281P+K309E, N59E+Y281P+K312Q,N59E+K308Q+K309E, N59E+K308Q+K312Q, N59E+K309E+K312Q, S163P+S186R+S225G,S163P+S186R+N227T, S163P+S186R+E232D, S163P+S186R+G235W,S163P+S186R+N252P, S163P+S186R+N260Q, S163P+S186R+H272V,S163P+S186R+S279D, S163P+S186R+Y281P, S163P+S186R+K308Q,S163P+S186R+K309E, S163P+S186R+K312Q, S163P+S225G+N227T,S163P+S225G+E232D, S163P+S225G+G235W, S163P+S225G+N252P,S163P+S225G+N260Q, S163P+S225G+H272V, S163P+S225G+S279D,S163P+S225G+Y281P, S163P+S225G+K308Q, S163P+S225G+K309E,S163P+S225G+K312Q, S163P+N227T+E232D, S163P+N227T+G235W,S163P+N227T+N252P, S163P+N227T+N260Q, S163P+N227T+H272V,S163P+N227T+S279D, S163P+N227T+Y281P, S163P+N227T+K308Q,S163P+N227T+K309E, S163P+N227T+K312Q, S163P+E232D+G235W,S163P+E232D+N252P, S163P+E232D+N260Q, S163P+E232D+H272V,S163P+E232D+S279D, S163P+E232D+Y281P, S163P+E232D+K308Q,S163P+E232D+K309E, S163P+E232D+K312Q, S163P+G235W+N252P,S163P+G235W+N260Q, S163P+G235W+H272V, S163P+G235W+S279D,S163P+G235W+Y281P, S163P+G235W+K308Q, S163P+G235W+K309E,S163P+G235W+K312Q, S163P+N252P+N260Q, S163P+N252P+H272V,S163P+N252P+S279D, S163P+N252P+Y281P, S163P+N252P+K308Q,S163P+N252P+K309E, S163P+N252P+K312Q, S163P+N260Q+H272V,S163P+N260Q+S279D, S163P+N260Q+Y281P, S163P+N260Q+K308Q,S163P+N260Q+K309E, S163P+N260Q+K312Q, S163P+H272V+S279D,S163P+H272V+Y281P, S163P+H272V+K308Q, S163P+H272V+K309E,S163P+H272V+K312Q, S163P+S279D+Y281P, S163P+S279D+K308Q,S163P+S279D+K309E, S163P+S279D+K312Q, S163P+Y281P+K308Q,S163P+Y281P+K309E, S163P+Y281P+K312Q, S163P+K308Q+K309E,S163P+K308Q+K312Q, S163P+K309E+K312Q, S186R+S225G+N227T,S186R+S225G+E232D, S186R+S225G+G235W, S186R+S225G+N252P,S186R+S225G+N260Q, S186R+S225G+H272V, S186R+S225G+S279D,S186R+S225G+Y281P, S186R+S225G+K308Q, S186R+S225G+K309E,S186R+S225G+K312Q, S186R+N227T+E232D, S186R+N227T+G235W,S186R+N227T+N252P, S186R+N227T+N260Q, S186R+N227T+H272V,S186R+N227T+S279D, S186R+N227T+Y281P, S186R+N227T+K308Q,S186R+N227T+K309E, S186R+N227T+K312Q, S186R+E232D+G235W,S186R+E232D+N252P, S186R+E232D+N260Q, S186R+E232D+H272V,S186R+E232D+S279D, S186R+E232D+Y281P, S186R+E232D+K308Q,S186R+E232D+K309E, S186R+E232D+K312Q, S186R+G235W+N252P,S186R+G235W+N260Q, S186R+G235W+H272V, S186R+G235W+S279D,S186R+G235W+Y281P, S186R+G235W+K308Q, S186R+G235W+K309E,S186R+G235W+K312Q, S186R+N252P+N260Q, S186R+N252P+H272V,S186R+N252P+S279D, S186R+N252P+Y281P, S186R+N252P+K308Q,S186R+N252P+K309E, S186R+N252P+K312Q, S186R+N260Q+H272V,S186R+N260Q+S279D, S186R+N260Q+Y281P, S186R+N260Q+K308Q,S186R+N260Q+K309E, S186R+N260Q+K312Q, S186R+H272V+S279D,S186R+H272V+Y281P, S186R+H272V+K308Q, S186R+H272V+K309E,S186R+H272V+K312Q, S186R+S279D+Y281P, S186R+S279D+K308Q,S186R+S279D+K309E, S186R+S279D+K312Q, S186R+Y281P+K308Q,S186R+Y281P+K309E, S186R+Y281P+K312Q, S186R+K308Q+K309E,S186R+K308Q+K312Q, S186R+K309E+K312Q, S225G+N227T+E232D,S225G+N227T+G235W, S225G+N227T+N252P, S225G+N227T+N260Q,S225G+N227T+H272V, S225G+N227T+S279D, S225G+N227T+Y281P,S225G+N227T+K308Q, S225G+N227T+K309E, S225G+N227T+K312Q,S225G+E232D+G235W, S225G+E232D+N252P, S225G+E232D+N260Q,S225G+E232D+H272V, S225G+E232D+S279D, S225G+E232D+Y281P,S225G+E232D+K308Q, S225G+E232D+K309E, S225G+E232D+K312Q,S225G+G235W+N252P, S225G+G235W+N260Q, S225G+G235W+H272V,S225G+G235W+S279D, S225G+G235W+Y281P, S225G+G235W+K308Q,S225G+G235W+K309E, S225G+G235W+K312Q, S225G+N252P+N260Q,S225G+N252P+H272V, S225G+N252P+S279D, S225G+N252P+Y281P,S225G+N252P+K308Q, S225G+N252P+K309E, S225G+N252P+K312Q,S225G+N260Q+H272V, S225G+N260Q+S279D, S225G+N260Q+Y281P,S225G+N260Q+K308Q, S225G+N260Q+K309E, S225G+N260Q+K312Q,S225G+H272V+S279D, S225G+H272V+Y281P, S225G+H272V+K308Q,S225G+H272V+K309E, S225G+H272V+K312Q, S225G+S279D+Y281P,S225G+S279D+K308Q, S225G+S279D+K309E, S225G+S279D+K312Q,S225G+Y281P+K308Q, S225G+Y281P+K309E, S225G+Y281P+K312Q,S225G+K308Q+K309E, S225G+K308Q+K312Q, S225G+K309E+K312Q,N227T+E232D+G235W, N227T+E232D+N252P, N227T+E232D+N260Q,N227T+E232D+H272V, N227T+E232D+S279D, N227T+E232D+Y281P,N227T+E232D+K308Q, N227T+E232D+K309E, N227T+E232D+K312Q,N227T+G235W+N252P, N227T+G235W+N260Q, N227T+G235W+H272V,N227T+G235W+S279D, N227T+G235W+Y281P, N227T+G235W+K308Q,N227T+G235W+K309E, N227T+G235W+K312Q, N227T+N252P+N260Q,N227T+N252P+H272V, N227T+N252P+S279D, N227T+N252P+Y281P,N227T+N252P+K308Q, N227T+N252P+K309E, N227T+N252P+K312Q,N227T+N260Q+H272V, N227T+N260Q+S279D, N227T+N260Q+Y281P,N227T+N260Q+K308Q, N227T+N260Q+K309E, N227T+N260Q+K312Q,N227T+H272V+S279D, N227T+H272V+Y281P, N227T+H272V+K308Q,N227T+H272V+K309E, N227T+H272V+K312Q, N227T+S279D+Y281P,N227T+S279D+K308Q, N227T+S279D+K309E, N227T+S279D+K312Q,N227T+Y281P+K308Q, N227T+Y281P+K309E, N227T+Y281P+K312Q,N227T+K308Q+K309E, N227T+K308Q+K312Q, N227T+K309E+K312Q, E232D+G235W+N252P, E232D+G235W+N260Q, E232D+G235W+H272V, E232D+G235W+S279D,E232D+G235W+Y281P, E232D+G235W+K308Q, E232D+G235W+K309E,E232D+G235W+K312Q, E232D+N252P+N260Q, E232D+N252P+H272V,E232D+N252P+S279D, E232D+N252P+Y281P, E232D+N252P+K308Q,E232D+N252P+K309E, E232D+N252P+K312Q, E232D+N260Q+H272V,E232D+N260Q+S279D, E232D+N260Q+Y281P, E232D+N260Q+K308Q,E232D+N260Q+K309E, E232D+N260Q+K312Q, E232D+H272V+S279D,E232D+H272V+Y281P, E232D+H272V+K308Q, E232D+H272V+K309E,E232D+H272V+K312Q, E232D+S279D+Y281P, E232D+S279D+K308Q,E232D+S279D+K309E, E232D+S279D+K312Q, E232D+Y281P+K308Q,E232D+Y281P+K309E, E232D+Y281P+K312Q, E232D+K308Q+K309E,E232D+K308Q+K312Q, E232 D+K309E+K312Q, G235W+N252P+N260Q,G235W+N252P+H272V, G235W+N252P+S279D, G235W+N252P+Y281P,G235W+N252P+K308Q, G235W+N252P+K309E, G235W+N252P+K312Q,G235W+N260Q+H272V, G235W+N260Q+S279D, G235W+N260Q+Y281P,G235W+N260Q+K308Q, G235W+N260Q+K309E, G235W+N260Q+K312Q,G235W+H272V+S279D, G235W+H272V+Y281P, G235W+H272V+K308Q,G235W+H272V+K309E, G235W+H272V+K312Q, G235W+S279D+Y281P,G235W+S279D+K308Q, G235W+S279D+K309E, G235W+S279D+K312Q,G235W+Y281P+K308Q, G235W+Y281P+K309E, G235W+Y281P+K312Q,G235W+K308Q+K309E, G235W+K308Q+K312Q, G235W+K309E+K312Q,N252P+N260Q+H272V, N252P+N260Q+S279D, N252P+N260Q+Y281P,N252P+N260Q+K308Q, N252P+N260Q+K309E, N252P+N260Q+K312Q,N252P+H272V+S279D, N252P+H272V+Y281P, N252P+H272V+K308Q,N252P+H272V+K309E, N252P+H272V+K312Q, N252P+S279D+Y281P,N252P+S279D+K308Q, N252P+S279D+K309E, N252P+S279D+K312Q,N252P+Y281P+K308Q, N252P+Y281P+K309E, N252P+Y281P+K312Q,N252P+K308Q+K309E, N252P+K308Q+K312Q, N252P+K309E+K312Q,N260Q+H272V+S279D, N260Q+H272V+Y281P, N260Q+H272V+K308Q,N260Q+H272V+K309E, N260Q+H272V+K312Q, N260Q+S279D+Y281P,N260Q+S279D+K308Q, N260Q+S279D+K309E, N260Q+S279D+K312Q,N260Q+Y281P+K308Q, N260Q+Y281P+K309E, N260Q+Y281P+K312Q,N260Q+K308Q+K309E, N260Q+K308Q+K312Q, N260Q+K309E+K312Q,H272V+S279D+Y281P, H272V+S279D+K308Q, H272V+S279D+K309E,H272V+S279D+K312Q, H272V+Y281P+K308Q, H272V+Y281P+K309E,H272V+Y281P+K312Q, H272V+K308Q+K309E, H272V+K308Q+K312Q,H272V+K309E+K312Q, S279D+Y281P+K308Q, S279D+Y281P+K309E,S279D+Y281P+K312Q, S279D+K308Q+K309E, S279D+K308Q+K312Q,S279D+K309E+K312Q, Y281P+K308Q+K309E, Y281P+K308Q+K312Q,Y281P+K309E+K312Q and K308Q+K309E+K312Q.

In one preferred embodiment the dispersin variant of the inventioncomprises at least three substitutions wherein the substitutions orsubstitution sets are selected from the group consisting of:

F276A+K308E+K309E, F276A+K308E+K312E, F276A+K309E+K312E,K308E+K309E+K312E, N227T+F276A+K309E, N227T+F276A+K312E,N227T+K308E+K309E, N227T+K308E+K312E, N227T+K309E+K312E,N227T+N252P+K308E, N227T+N252P+K309E, N227T+N252P+K312E,N227T+N252P+N267T, N227T+N267T+F276A, N227T+N267T+K308E,N227T+N267T+K309E, N227T+N267T+K312E, N252P+F276A+K308E,N252P+F276A+K309E, N252P+F276A+K312E, N252P+K308E+K309E,N252P+K308E+K312E, N252P+N267T+F276A, N252P+N267T+K308E,N252P+N267T+K309E, N252P+N267T+K312E, N267T+F276A+K308E,N267T+F276A+K309E, N267T+F276A+K312E, N267T+K308E+K309E,N267T+K308E+K312E, N267T+K309E+K312E, Q215K+F276A+K308E,Q215K+F276A+K312E, Q215K+K308E+K312E, Q215K+K309E+K312E,Q215K+N227T+F276A, Q215K+N227T+K308E, Q215K+N227T+K309E,Q215K+N227T+K312E, Q215K+N227T+N252P, Q215K+N227T+N267T,Q215K+N252P+F276A, Q215K+N252P+K309E, Q215K+N252P+K312E,Q215K+N252P+N267T, Q215K+N267T+F276A, Q215K+N267T+K312E,S163P+F276A+K308E, S163P+F276A+K312E, S163P+K308E+K309E,S163P+K308E+K312E, S163P+K309E+K312E, S163P+N227T+F276A,S163P+N227T+K309E, S163P+N227T+N252P, S163P+N227T+N267T,S163P+N252P+F276A, S163P+N252P+K308E, S163P+N252P+K309E,S163P+N252P+K312E, S163P+N252P+N267T, S163P+N267T+K308E,S163P+N267T+K309E, S163P+N267T+K312E, S163P+Q215K+F276A,S163P+Q215K+K308E, S163P+Q215K+K309E, S163P+Q215K+K312E,S163P+Q215K+N227T and S163P+Q215K+N252P.

In one preferred embodiment the dispersin variant of the inventioncomprises at least four substitutions wherein the substitutions orsubstitution sets are selected from the group consisting of:K312E+N227T+F276A+K308E, K312E+N227T+F276A+K309E,K312E+N227T+F276A+K312E, K312E+N227T+K308E+K309E,K312E+N227T+K308E+K312E, K312E+N227T+K309E+K312E,K312E+F276A+K308E+K309E, K312E+F276A+K308E+K312E,K312E+F276A+K309E+K312E, K312E+K308E+K309E+K312E,N227T+F276A+K308E+K309E, N227T+F276A+K308E+K312E,N227T+F276A+K309E+K312E, N227T+K308E+K309E+K312E,F276A+K308E+K309E+K312E, F276A+K308E+K309E+K312E,N227T+K308E+K309E+K312E and N227T+F276A+K308E+K309E.

In one preferred embodiment the dispersin variant of the inventioncomprises at least four substitutions wherein the substitution sets areselected from the group consisting of: K312Q+N227T+S225G+K308Q,K312Q+N227T+S225G+K309E, K312Q+N227T+S225G+K312Q,K312Q+N227T+K308Q+K309E, K312Q+N227T+K308Q+K312Q,K312Q+N227T+K309E+K312Q, K312Q+S225G+K308Q+K309E,K312Q+S225G+K308Q+K312Q, K312Q+S225G+K309E+K312Q,K312Q+K308Q+K309E+K312Q, N227T+S225G+K308Q+K309E,N227T+S225G+K308Q+K312Q, N227T+S225G+K309E+K312Q,N227T+K308Q+K309E+K312Q, S225G+K308Q+K309E+K312Q,S225G+K308Q+K309E+K312Q, N227T+K308Q+K309E+K312Q andN227T+S225G+K308Q+K309E.

In one preferred embodiment the dispersin variant of the inventioncomprises at least four substitutions wherein the substitutions orsubstitution sets are selected from the group consisting of:N227T+F276A+K308E+K309E, N227T+F276A+K308E+K312E,N227T+F276A+K309E+K312E, N227T+N267T+F276A+K308E,N227T+N267T+F276A+K309E, N227T+N267T+F276A+K312E, andS163P+N227T+F276A+K308E,

In one preferred embodiment the dispersin variant of the inventioncomprises at least four substitutions wherein the substitution sets areselected from the group consisting of: N227T+S225G+K308Q+K309E,N227T+S225G+K308Q+K312Q, N227T+S225G+K309E+K312Q,N227T+N267T+S225G+K308Q, N227T+N267T+S225G+K309E,N227T+N267T+S225G+K312Q, and S163P+N227T+S225G+K308Q,

In one preferred embodiment the dispersin variant of the inventioncomprises at least five substitutions wherein the substitutions orsubstitution sets are selected from the group consisting of:S163P+Q215K+N227T+N252P+N267T, S163P+Q215K+N227T+N252P+F276A,S163P+Q215K+N227T+N252P+K308E, S163P+Q215K+N227T+N252P+K309E,S163P+Q215K+N227T+N267T+F276A, S163P+Q215K+N227T+N267T+K308E,S163P+Q215K+N227T+N267T+K309E, S163P+Q215K+N227T+F276A+K308E,S163P+Q215K+N227T+F276A+K309E, S163P+Q215K+N227T+K308E+K309E,S163P+Q215K+N252P+N267T+F276A, S163P+Q215K+N252P+N267T+K308E,S163P+Q215K+N252P+N267T+K309E, S163P+Q215K+N252P+F276A+K308E,S163P+Q215K+N252P+F276A+K309E, S163P+Q215K+N252P+K308E+K309E,S163P+Q215K+N267T+F276A+K308E, S163P+Q215K+N267T+F276A+K309E,S163P+Q215K+N267T+K308E+K309E, S163P+Q215K+F276A+K308E+K309E,S163P+N227T+N252P+N267T+F276A, S163P+N227T+N252P+N267T+K308E,S163P+N227T+N252P+N267T+K309E, S163P+N227T+N252P+F276A+K308E,S163P+N227T+N252P+F276A+K309E, S163P+N227T+N252P+K308E+K309E,S163P+N227T+N267T+F276A+K308E, S163P+N227T+N267T+F276A+K309E,S163P+N227T+N267T+K308E+K309E, S163P+N227T+F276A+K308E+K309E,S163P+N252P+N267T+F276A+K308E, S163P+N252P+N267T+F276A+K309E,S163P+N252P+N267T+K308E+K309E, S163P+N252P+F276A+K308E+K309E,S163P+N267T+F276A+K308E+K309E, Q215K+N227T+N252P+N267T+F276A,Q215K+N227T+N252P+N267T+K308E, Q215K+N227T+N252P+N267T+K309E,Q215K+N227T+N252P+F276A+K308E, Q215K+N227T+N252P+F276A+K309E,Q215K+N227T+N252P+K308E+K309E, Q215K+N227T+N267T+F276A+K308E,Q215K+N227T+N267T+F276A+K309E, Q215K+N227T+N267T+K308E+K309E,Q215K+N227T+F276A+K308E+K309E, Q215K+N252P+N267T+F276A+K308E,Q215K+N252P+N267T+F276A+K309E, Q215K+N252P+N267T+K308E+K309E,Q215K+N252P+F276A+K308E+K309E, Q215K+N267T+F276A+K308E+K309E,N227T+N252P+N267T+F276A+K308E, N227T+N252P+N267T+F276A+K309E,N227T+N252P+N267T+K308E+K309E, N227T+N252P+F276A+K308E+K309E,N227T+N267T+F276A+K308E+K309E N252P+N267T+F276A+K308E+K309E,Q215K+N227T+F276A+K308E+K309E, Q215K+N227T+F276A+K308E+K312E,Q215K+N227T+F276A+K309E+K312E, Q215K+N227T+K308E+K309E+K312E,Q215K+F276A+K308E+K309E+K312E, and N227T+F276A+K308E+K309E+K312E.

In one preferred embodiment the dispersin variant of the inventioncomprises at least five substitutions wherein the substitution sets areselected from the group consisting of: S163P+N59E+N227T+N252P+E232D,S163P+N59E+N227T+N252P+S225G, S163P+N59E+N227T+N252P+K308Q,S163P+N59E+N227T+N252P+K309E, S163P+N59E+N227T+E232D+S225G,S163P+N59E+N227T+E232D+K308Q, S163P+N59E+N227T+E232D+K309E,S163P+N59E+N227T+S225G+K308Q, S163P+N59E+N227T+S225G+K309E,S163P+N59E+N227T+K308Q+K309E, S163P+N59E+N252P+E232D+S225G,S163P+N59E+N252P+E232D+K308Q, S163P+N59E+N252P+E232D+K309E,S163P+N59E+N252P+S225G+K308Q, S163P+N59E+N252P+S225G+K309E,S163P+N59E+N252P+K308Q+K309E, S163P+N59E+E232 D+S225G+K308Q,S163P+N59E+E232D+S225G+K309E, S163P+N59E+E232 D+K308Q+K309E,S163P+N59E+S225G+K308Q+K309E, S163P+N227T+N252P+E232D+S225G,S163P+N227T+N252P+E232D+K308Q, S163P+N227T+N252P+E232D+K309E,S163P+N227T+N252P+S225G+K308Q, S163P+N227T+N252P+S225G+K309E,S163P+N227T+N252P+K308Q+K309E, S163P+N227T+E232D+S225G+K308Q,S163P+N227T+E232D+S225G+K309E, S163P+N227T+E232D+K308Q+K309E,S163P+N227T+S225G+K308Q+K309E, S163P+N252P+E232D+S225G+K308Q,S163P+N252P+E232 D+S225G+K309E, S163P+N252P+E232D+K308Q+K309E,S163P+N252P+S225G+K308Q+K309E, S163P+E232D+S225G+K308Q+K309E,N59E+N227T+N252P+E232D+S225G, N59E+N227T+N252P+E232D+K308Q,N59E+N227T+N252P+E232D+K309E, N59E+N227T+N252P+S225G+K308Q,N59E+N227T+N252P+S225G+K309E, N59E+N227T+N252P+K308Q+K309E,N59E+N227T+E232D+S225G+K308Q, N59E+N227T+E232 D+S225G+K309E,N59E+N227T+E232D+K308Q+K309E, N59E+N227T+S225G+K308Q+K309E,N59E+N252P+E232D+S225G+K308Q, N59E+N252P+E232D+S225G+K309E,N59E+N252P+E232D+K308Q+K309E, N59E+N252P+S225G+K308Q+K309E,N59E+E232D+S225G+K308Q+K309E, N227T+N252P+E232D+S225G+K308Q,N227T+N252P+E232D+S225G+K309E, N227T+N252P+E232D+K308Q+K309E,N227T+N252P+S225G+K308Q+K309E, N227T+E232D+S225G+K308Q+K309EN252P+E232D+S225G+K308Q+K309E, N59E+N227T+S225G+K308Q+K309E,N59E+N227T+S225G+K308Q+K312Q, N59E+N227T+S225G+K309E+K312Q,N59E+N227T+K308Q+K309E+K312Q, N59E+S225G+K308Q+K309E+K312Q, andN227T+S225G+K308Q+K309E+K312Q.

In one preferred embodiment the dispersin variant of the inventioncomprises at least five substitutions wherein the substitutions orsubstitution sets are selected from the group consisting of:N227T+N267T+F276A+K308E+K309E, N227T+N267T+F276A+K308E+K312E,N227T+N267T+F276A+K309E+K312E, S163P+N227T+F276A+K308E+K312E,N227T+F276A+K308E+K309E+K312E, N227T+F276A+K308E+K309E+K312E, andN227T+F276A+K308E+K309E+K312E.

In one preferred embodiment the dispersin variant of the inventioncomprises at least five substitutions wherein the substitution sets areselected from the group consisting of:

N227T+E232D+S225G+K308Q+K309E, N227T+E232D+S225G+K308Q+K312Q,N227T+E232D+S225G+K309E+K312Q, S163P+N227T+S225G+K308Q+K312Q,N227T+S225G+K308Q+K309E+K312Q, N227T+S225G+K308Q+K309E+K312Q, andN227T+S225G+K308Q+K309E+K312Q.

In one preferred embodiment the dispersin variant of the inventioncomprises at least six substitutions wherein the substitutions orsubstitution sets are selected from the group consisting of:

N227T+N267T+F276A+K308E+K309E+K312E,N227T+N267T+F276A+K308E+K309E+K312E,N227T+N252P+F276A+K308E+K309E+K312E,Q215K+N227T+F276A+K308E+K309E+K312E,S163P+N227T+F276A+K308E+K309E+K312E,N227T+N267T+F276A+K308E+K309E+K312E,N227T+N252P+F276A+K308E+K309E+K312E, Q215K+N227T+F276A+K308E+K309E+K312Eand S163P+N227T+F276A+K308E+K309E+K312E.

In one preferred embodiment the dispersin variant of the inventioncomprises at least six substitutions wherein the substitution sets areselected from the group consisting of:

N227T+S225G+S186R+K308Q+K309E+K312Q,N227T+S225G+S186R+K308Q+K309E+K312Q,N227T+N252P+S186R+K308Q+K309E+K312Q,G235W+N227T+S186R+K308Q+K309E+K312Q,S163P+N227T+S186R+K308Q+K309E+K312Q,N227T+S225G+S186R+K308Q+K309E+K312Q,N227T+N252P+S186R+K308Q+K309E+K312Q, G235W+N227T+S186R+K308Q+K309E+K312Qand S163P+N227T+S186R+K308Q+K309E+K312Q.

In one preferred embodiment the dispersin variant of the inventioncomprises at least seven substitutions wherein the substitutions orsubstitution sets are selected from the group consisting of:N227T+N252P+N267T+F276A+K308E+K309E+K312E,Q215K+N227T+N252P+F276A+K308E+K309E+K312E,Q215K+N227T+N267T+F276A+K308E+K309E+K312E,S163P+N227T+N252P+F276A+K308E+K309E+K312E,S163P+N227T+N267T+F276A+K308E+K309E+K312E,S163P+Q215K+N227T+F276A+K308E+K309E+K312E,N227T+N252P+N267T+F276A+K308E+K309E+K312E,Q215K+N227T+N252P+F276A+K308E+K309E+K312E,Q215K+N227T+N267T+F276A+K308E+K309E+K312E,S163P+N227T+N252P+F276A+K308E+K309E+K312E,S163P+N227T+N267T+F276A+K308E+K309E+K312E, andS163P+Q215K+N227T+F276A+K308E+K309E+K312E.

In one preferred embodiment the dispersin variant of the inventioncomprises at least seven substitutions wherein the substitution sets areselected from the group consisting of:N227T+N252P+S225G+S186R+K308Q+K309E+K312Q,S186R+N227T+N252P+S186R+K308Q+K309E+K312Q,S186R+N227T+S225G+S186R+K308Q+K309E+K312Q,S163P+N227T+N252P+S186R+K308Q+K309E+K312Q,S163P+N227T+S225G+S186R+K308Q+K309E+K312Q,S163P+S186R+N227T+S186R+K308Q+K309E+K312Q,N227T+N252P+S225G+S186R+K308Q+K309E+K312Q,S186R+N227T+N252P+S186R+K308Q+K309E+K312Q,S186R+N227T+S225G+S186R+K308Q+K309E+K312Q,S163P+N227T+N252P+S186R+K308Q+K309E+K312Q,S163P+N227T+S225G+S186R+K308Q+K309E+K312Q, andS163P+S186R+N227T+S186R+K308Q+K309E+K312Q.

In one preferred embodiment the dispersin variant of the inventioncomprises at least eight substitutions wherein the substitutions areselected from the group consisting of:S163P+Q215K+N227T+N252P+N267T+F276A+K308E+K309E,S163P+Q215K+N227T+N252P+N267T+F276A+K308E+K312E,S163P+Q215K+N227T+N252P+N267T+F276A+K309E+K312E,S163P+Q215K+N227T+N252P+N267T+K308E+K309E+K312E,S163P+Q215K+N227T+N252P+F276A+K308E+K309E+K312E,S163P+Q215K+N227T+N267T+F276A+K308E+K309E+K312E,S163P+Q215K+N252P+N267T+F276A+K308E+K309E+K312E,S163P+N227T+N252P+N267T+F276A+K308E+K309E+K312E, andQ215K+N227T+N252P+N267T+F276A+K308E+K309E+K312E.

In one preferred embodiment the dispersin variant of the inventioncomprises at least eight substitutions wherein the substitution sets areselected from the group consisting of:S163P+G235W+N227T+N252P+S186R+N260Q+K308Q+K309E,S163P+G235W+N227T+N252P+S186R+N260Q+K308Q+K312Q,S163P+G235W+N227T+N252P+S186R+N260Q+K309E+K312Q,S163P+G235W+N227T+N252P+S186R+K308Q+K309E+K312Q,S163P+G235W+N227T+N252P+N260Q+K308Q+K309E+K312Q,S163P+G235W+N227T+S186R+N260Q+K308Q+K309E+K312Q,S163P+G235W+N252P+S186R+N260Q+K308Q+K309E+K312Q,S163P+N227T+N252P+S186R+N260Q+K308Q+K309E+K312Q, andG235W+N227T+N252P+S186R+N260Q+K308Q+K309E+K312Q.

In one preferred embodiment the dispersin variant of the inventioncomprises at least eight substitutions wherein the substitutions orsubstitution sets are selected from the group consisting of:Q215K+N227T+N252P+N267T+F276A+K308E+K309E+K312E,S163P+N227T+N252P+N267T+F276A+K308E+K309E+K312E,S163P+Q215K+N227T+N252P+F276A+K308E+K309E+K312E,S163P+Q215K+N227T+N267T+F276A+K308E+K309E+K312E,Q215K+N227T+N252P+N267T+F276A+K308E+K309E+K312E,S163P+N227T+N252P+N267T+F276A+K308E+K309E+K312E,S163P+Q215K+N227T+N252P+F276A+K308E+K309E+K312E andS163P+Q215K+N227T+N267T+F276A+K308E+K309E+K312E.

In one preferred embodiment the dispersin variant of the inventioncomprises at least eight substitutions wherein the substitution sets areselected from the group consisting of:S225G+N227T+N252P+S186R+N260Q+K308E+K309E+K312E,S163P+N227T+N252P+S186R+N260Q+K308E+K309E+K312E,S163P+S225G+N227T+N252P+N260Q+K308E+K309E+K312E,S163P+S225G+N227T+S186R+N260Q+K308E+K309E+K312E,S225G+N227T+N252P+S186R+N260Q+K308E+K309E+K312E,S163P+N227T+N252P+S186R+N260Q+K308E+K309E+K312E,S163P+S225G+N227T+N252P+N260Q+K308E+K309E+K312E andS163P+S225G+N227T+S186R+N260Q+K308E+K309E+K312E.

In one preferred embodiment the dispersin variant of the inventioncomprises at least nine substitutions wherein the substitutions orsubstitution sets are selected from the group consisting of:S163P+Q215K+N227T+N252P+N267T+F276A+K308E+K309E+K312E,Q3F+Q215K+N227T+N252P+N267T+F276A+K308E+K309E+K312E,A49W+S163P+Q215K+N227T+N252P+N267T+F276A+K309E+K312E,Q3F+S163P+Q215K+N227T+N252P+N267T+K308E+K309E+K312E, andA49W+Q215K+N227T+N252P+N267T+F276A+K308E+K309E+K312E.

In one preferred embodiment the dispersin variant of the inventioncomprises at least nine substitutions wherein the substitution sets areselected from the group consisting of:S163P+S225G+N227T+N252P+N260Q+S186R+K308Q+K309E+K312Q,Q3I+S225G+N227T+N252P+N260Q+S186R+K308Q+K309E+K312Q,A49W+S163P+S225G+N227T+N252P+N260Q+S186R+K309E+K312Q,Q3I+S163P+S225G+N227T+N252P+N260Q+K308Q+K309E+K312Q, andA49W+S225G+N227T+N252P+N260Q+S186R+K308Q+K309E+K312Q.

In one preferred embodiment the dispersin variant of the inventioncomprises at least nine substitutions wherein the substitutions orsubstitution sets are selected from the group consisting of:S163P+Q215K+N227T+N252P+N267T+F276A+K308E+K309E+K312E andS163P+Q215K+N227T+N252P+N267T+F276A+K308E+K309E+K312E.

In one preferred embodiment the dispersin variant of the inventioncomprises at least nine substitutions wherein the substitution sets areselected from the group consisting of:S163P+S186R+S225G+N227T+G235W+N252P+K308Q+K309E+K312Q andS163P+S186R+S225G+N227T+G235W+N252P+K308Q+K309E+K312Q.

In one preferred embodiment the dispersin variant of the inventioncomprises at least eight, at least nine, at least ten or at least elevensubstitutions wherein the substitutions or substitution sets areselected from the group consisting of:Q3F+A49W+Q215K+F276A+K308E+K309E+K312E+N227T+N267T,Q3F+A49W+Q215K+F276A+K308E+K309E+K312E+N227T+N267T,Q3F+A49W+Q215K+F276A+K308E+K309E+K312E+N227T+N267T,Q3F+A49W+Q215K+F276A+K308E+K309E+K312E+N227T+,Q3F+A49W+Q215K+F276A+K308E+K309E+K312E+N267T+,Q3F+A49W+Q215K+F276A+K308E+K309E+N227T+N267T+,Q3F+A49W+Q215K+F276A+K308E+K312E+N227T+N267T+,Q3F+A49W+Q215K+F276A+K309E+K312E+N227T+N267T+,Q3F+A49W+Q215K+K308E+K309E+K312E+N227T+N267T+,Q3F+A49W+F276A+K308E+K309E+K312E+N227T+N267T+,Q3F+Q215K+F276A+K308E+K309E+K312E+N227T+N267T+,A49W+Q215K+F276A+K308E+K309E+K312E+N227T+N267T+andQ3F+A49W+Q215K+F276A+K308E+K309E+K312E+N227T+N267T.

In one preferred embodiment the dispersin variant of the inventioncomprises at least eleven substitutions wherein the substitution setsare selected from the group consisting of:Q3I+A49W+N59E+S163P+S186R+S225G+N227T+Y281P+K308Q+K309E+K312Q,Q3I+A49W+N59E+S163P+S186R+S225G+E232D+Y281P+K308Q+K309E+K312Q,Q3I+A49W+N59E+S163P+S186R+S225G+G235W+Y281P+K308Q+K309E+K312Q,Q3I+A49W+N59E+S163P+S186R+S225G+N252P+Y281P+K308Q+K309E+K312Q,Q3I+A49W+N59E+S163P+S186R+S225G+N260Q+Y281P+K308Q+K309E+K312Q,Q3I+A49W+N59E+S163P+S186R+S225G+N227T+H272V+Y281P+K308Q+K309E,Q3I+A49W+N59E+S163P+S186R+S225G+N227T+S279D+Y281P+K308Q+K312Q,H15Y+A49W+N59E+S163P+S186R+S225G+N227T+E232 D+Y281P+K309E+K312Q,H15Y+A49W+N 59E+S163P+S186R+S225G+N227T+G235W+K308Q+K309E+K312Q,H15Y+A49W+S163P+S186R+S225G+N227T+Y281P+S288P+K308Q+K309E+K312Q,H15Y+A49W+S163P+S186R+S225G+N227T+N252P+Y281P+K308Q+K309E+K312Q,H15Y+A49W+S163P+S186R+S225G+N227T+H272P+Y281P+K308Q+K309E+K312Q,H15Y+A49W+S163P+S186R+S225G+N227T+S279D+Y281P+K308Q+K309E+K312Q, andH15Y+A49W+N59E+S163P+S186R+S225G+N227T+E232D+Y281P+K308Q+K312Q.

In one preferred embodiment of the invention the dispersin variant ofthe invention comprises one of the substitution sets selected from thegroup consisting of:

Q3F+A49W+N59E+S163P+S186R+S225G+N227T+N252P+F276A+Y281P+K308E+K309E+K312E;

Q3I+H15Y+A49W+N59E+S163P+S186R+S225G+N227T+E232D+N252P+N260Q+N267T+H272V+S279D+Y281P+K308Q+K309E+K312Q;

Q3I+H15Y+A49W+N59E+S163P+S186R+S225G+N227T+E232D+N252P+N260Q+N267T+H272P+S279D+Y281P+K308Q+K309E+K312Q;

Q3I+H15Y+A49W+N59E+S163P+S186R+S225G+N227T+E232D+G235W+N252P+N260Q+H272V+S279D+Y281P+K308Q+K309E+K312E;

Q3I+H15Y+A49W+N59E+S163P+S186R+S225G+N227T+E232D+G235W+N252P+N260Q+H272V+S279D+Y281P+K308E+K309E+K312Q;

Q3F+H15Y+V1401+S163P+S186R+Q215K+S225G+N227T+E232D+G235W+N252P+N260Q+N267T+H272V+S279D+Y281P+K308Q+K309E+K312Q;Q3F+H15Y+S163P+S186R+Q215K+S225G+N227T+E232D+N252P+N260Q+N267T+H272V+S279D+Y281P+K308Q+K309E+K312Q;Q3F+H15Y+T17W+A49W+N59E+S163P+S186R+T218Q+S225G+N227T+G235W+S237W+N252P+H272P+K308Q+K309E+K312Q;H15Y+A49W+N59E+S163P+S186R+S225G+N227T+E232D+G235W+N252P+N260Q+H272V+S279D+Y281P+K309E+K312Q;Q3F+H15Y+T17W+A49W+N59E+S163P+S186R+D207N+T218Q+S225G+N227T+G235W+S237W+N252P+H272P+Y281P+K308Q+K309E+K312Q;Q3I+H15Y+A49W+N59E+S163P+S186R+S225G+N227T+E232D+G235W+N252P+N260Q+H272V+S279D+Y281P+K308Q+K309E+K312Q;H15Y+A49W+N59E+S163P+S186R+S225G+N227T+E232D+N252P+N260Q+H272P+S279D+Y281P+S288P+K308Q+K309E+K312Q;andH15Y+A49W+N59E+S163P+S186R+S225G+N227T+E232D+N252P+N260Q+H272P+S279D+Y281P+K308Q+K309E+K312Q,wherein the variant has at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 95%, suchas at least 96%, at least 97%, at least 98%, or at least 99%, but lessthan 100% sequence identity to the polypeptide shown in SEQ ID NO: 1,wherein each position corresponds to the position of the polypeptideshown in SEQ ID NO: 1 (numbering according to SEQ ID NO: 1).

In one preferred embodiment of the invention the dispersin variant ofthe invention comprises one of the substitution sets selected from thegroup consisting of: S186R+N227T+E232D+K308E,S163P+Q215K+N227T+N252P+N267T+F276A+K308Q+K309E+K312E,S163P+Q215K+N227T+N252P+N267T+F276A+K308E+K309E+K312Q,

Q3F+A49W+N59E+S163P+S186R+N227T+E232D+N252P+F276A+S279G+K308E+K309E+K312E,

A49W+N59E+S163P+S186R+S225G+N227T+E232D+N252P+F276A+S279G+K308E+K309E+K312E,

Q3F+T17W+A49W+N59E+S163P+S186R+S225G+N227T+G235W+N252P+H272P+Y281P+K308Q+K309E+K312Q,

A49W+N59E+S163P+S186R+N227T+E232D+N252P+H272V+F276A+S279G+K308E+K309E+K312E,

Q3F+A49W+N59E+S163P+S186R+S225G+N227T+N252P+F276A+S279G+K308E+K309E+K312E,

A49W+N59E+S163P+S186R+N227T+E232D+N252P+F276A+S279G+Y281P+K308E+K309E+K312E,

A49W+N59E+S163P+S186R+S225G+N227T+N252P+F276A+S279G+Y281P+K308E+K309E+K312E,

Q3F+A49W+N59E+S163P+S186R+S225G+N227T+E232D+N252P+F276A+S279G+K308E+K309E+K312E,

Q3F+S163P+S186R+Q215K+N227T+E232D+N252P+N267T+F276A+S279G+K308E+K309E+K312E,

H15Y+A49W+N59E+S163P+S186R+S225G+N227T+E232D+G235W+N252P+N260Q+H272P+S279D+Y281P+K308Q+K309E+K312Q,

Q3F+S163P+S186R+Q215K+S225G+N227T+N252P+N267T+F276A+S279G+K308E+K309E+K312E,

H15Y+A49W+N59E+S163P+S186R+S225G+N227T+E232D+N252P+N260Q+H272P+S279D+Y281P+S288P+K308Q+K309E+K312Q,

A49W+N59E+S163P+S186R+S225G+N227T+E232D+N252P+H272V+F276A+S279G+K308E+K309E+K312E,

H15Y+A49W+N59E+S163P+S186R+S225G+N227T+E232D+G235W+N252P+N260Q+H272V+S279D+Y281P+K308Q+K309E+K312Q,

S163P+S186R+Q215K+N227T+E232D+N252P+N267T+F276A+S279G+Y281P+K308E+K309E+K312E,

Q3F+A49W+N59E+S163P+S186R+N227T+E232D+N252P+F276A+S279G+Y281P+K308E+K309E+K312E,

H15Y+A49W+N59E+S163P+S186R+S225G+N227T+E232D+N252P+N260Q+H272P+S279D+Y281P+K308Q+K309E+K312Q,

A49W+N59E+S163P+S186R+S225G+N227T+E232D+N252P+F276A+S279G+Y281P+K308E+K309E+K312E,

H15Y+A49W+N59E+S163P+S186R+S225G+N227T+E232D+N252P+N260Q+H272V+Y281P+K308Q+K309E+K312Q,

Q3F+T17W+A49W+N59E+S163P+S186R+D207N+T218Q+S225G+N227T+G235W+N252P+H272P+Y281P+K308Q+K309E+K312Q,

H15Y+T17W+A49W+N59E+S163P+S186R+S225G+N227T+E232D+G235W+N252P+N260Q+H272V+S279D+Y281P+K308Q+K309E+K312Q,

Q3F+A49W+N59E+S163P+S186R+S225G+N227T+N252P+F276A+S279G+Y281P+K308E+K309E+K312E,

Q3F+A49W+N59E+S163P+S186R+S225G+N227T+E232D+N252P+H272V+F276A+S279G+K308E+K309 E+K312E,

A49W+N59E+S163P+S186R+N227T+E232D+N252P+H272V+F276A+S279G+Y281P+K308E+K309E+K312E,

S163P+S186R+S225G+N227T+E232D+N252P+N267T+F276A+S279G+Y281P+K308E+K309E+K312E,

H15Y+T17W+A49W+N59E+S163P+S186R+S225G+N227T+E232D+N252P+N260Q+H272V+S279D+Y281P+K308Q+K309E+K312Q,

Q3F+S163P+S186R+Q215K+N227T+E232D+N252P+N267T+H272V+F276A+S279G+K308E+K309E+K312E,

Q3I+H15Y+A49W+N59E+S163P+S186R+S225G+N227T+E232D+G235W+N252P+N260Q+H272V+S279D+Y281P+K308Q+K309E+K312Q,

S163P+S186R+Q215K+S225G+N227T+E232D+N252P+N267T+F276A+S279G+Y281P+K308E+K309E+K312E,

S163P+Q215K+S225G+N227T+E232D+N252P+N267T+F276A+S279G+Y281P+K308E+K309E+K312E,

S163P+S186R+Q215K+S225G+N227T+E232D+N252P+N267T+H272V+F276A+S279G+K308E+K309E+K312E,

S163P+Q215K+S225G+N227T+E232D+N252P+N267T+H272V+F276A+S279G+K308E+K309E+K312E,

A49W+N59E+S163P+S186R+S225G+N227T+N252P+H272V+F276A+S279G+Y281P+K308E+K309E+K312E,

Q3F+S163P+S186R+Q215K+N227T+E232D+N252P+N267T+F276A+S279G+Y281P+K308E+K309E+K312E,

Q3F+S163P+Q215K+N227T+E232D+N252P+N267T+F276A+S279G+Y281P+K308E+K309E+K312E,

Q3F+H15Y+T17W+A49W+N59E+S163P+S186R+T218Q+S225G+N227T+G235W+S237W+N252P+H272P+Y281P+K308Q+K309E+K312Q,

S163P+Q215K+N227T+E232D+N252P+N267T+H272V+F276A+S279G+Y281P+K308E+K309E+K312E,

Q3F+S163P+Q215K+S225G+N227T+E232D+N252P+N267T+H272V+F276A+S279G+K308E+K309E+K312E,

Q3F+A49W+N59E+S163P+S186R+N227T+E232D+N252P+H272V+F276A+S279G+Y281P+K308E+K309E+K312E,

Q3F+S163P+S186R+Q215K+S225G+N227T+N252P+N267T+F276A+S279G+Y281P+K308E+K309E+K312E,

Q3F+H15Y+T17W+A49W+N59E+S163P+S186R+D207N+T218Q+S225G+N227T+G235W+S237W+N252P+H272P+Y281P+K308Q+K309E+K312Q,

S163P+S186R+Q215K+S225G+N227T+N252P+N267T+H272V+F276A+S279G+Y281P+K308E+K309E+K312E,

S163P+Q215K+S225G+N227T+N252P+N267T+H272V+F276A+S279G+Y281P+K308E+K309E+K312E,

Q3F+S163P+Q215K+N227T+E232D+N252P+N267T+H272V+F276A+S279G+Y281P+K308E+K309E+K312E,

Q3F+A49W+N59E+S163P+S186R+S225G+N227T+E232D+N252P+F276A+S279G+Y281P+K308E+K309E+K312E,

Q3F+A49W+N59E+S163P+S186R+S225G+N227T+N252P+H272V+F276A+S279G+Y281P+K308E+K309E+K312E,

A49W+N59E+S163P+S186R+S225G+N227T+E232D+N252P+H272V+F276A+S279G+Y281P+K308E+K309E+K312E,

S163P+Q215K+S225G+N227T+E232D+N252P+N267T+H272V+F276A+S279G+Y281P+K308E+K309E+K312E,

Q3F+S163P+S186R+Q215K+S225G+N227T+E232D+N252P+N260Q+N267T+H272P+S279D+Y281P+K308Q+K309E+K312Q,

Q3F+S163P+S186R+Q215K+N227T+E232D+N252P+N267T+H272V+F276A+S279G+Y281P+K308E+K309E+K312E,

S163P+S186R+Q215K+S225G+N227T+E232D+N252P+N267T+H272V+F276A+S279G+Y281P+K308E+K309E+K312E,

Q3F+S163P+S186R+Q215K+S225G+N227T+N252P+N267T+H272V+F276A+S279G+Y281P+K308E+K309E+K312E,

Q3F+H15Y+V1401+S163P+S186R+Q215K+S225G+N227T+E232D+G235W+N252P+N260Q+N267T+H272V+S279D+Y281P+K308Q+K309E+K312Q,

Q3F+T17W+S163P+S186R+Q215K+S225G+N227T+E232D+G235W+N252P+N267T+H272V+F276A+S279G+Y281P+K308E+K309E+K312Q,

Q3F+S163P+S186R+Q215K+S225G+N227T+E232D+N252P+N267T+H272V+F276A+S279G+Y281P+K308E+K309E+K312Q,

Q3F+A49W+N59E+S163P+S186R+S225G+N227T+E232D+N252P+H272V+F276A+S279G+Y281P+K308E+K309E+K312E,

Q3F+S163P+S186R+Q215K+S225G+N227T+E232D+N252P+N267T+H272V+F276A+S279G+Y281P+K308E+K309E+K312E,

Q3F+H15Y+T17W+S163P+S186R+Q215K+S225G+N227T+E232D+G235W+N252P+N260Q+N267T+H272V+S279D+Y281P+K308Q+K309E+K312Q,

Q3F+S163P+Q215K+S225G+N227T+E232D+N252P+N267T+H272V+F276A+S279G+Y281P+K308E+K309E+K312E,

Q3F+H15Y+S163P+S186R+Q215K+S225G+N227T+E232D+G235W+N252P+N260Q+N267T+H272P+S279D+Y281P+K308Q+K309E+K312Q,

Q3F+A49W+N59E+S163P+S186R+S225G+N227T+E232D+G235W+N252P+N260Q+H272P+F276A+S279D+Y281P+K308E+K309E+K312Q,

Q3F+H15Y+T17W+S163P+S186R+Q215K+S225G+N227T+E232D+N252P+N260Q+N267T+H272V+S279D+Y281P+K308Q+K309E+K312Q,

Q3F+H15Y+S163P+S186R+Q215K+S225G+N227T+E232D+N252P+N260Q+N267T+H272P+S279D+Y281P+K308Q+K309E+K312Q,

Q3F+H15Y+S163P+S186R+Q215K+S225G+N227T+E232D+G235W+N252P+N267T+H272V+F276A+S279G+Y281P+K308Q+K309E+K312Q,

Q3F+H15Y+T17W+S163P+S186R+Q215K+S225G+N227T+E232D+N252P+N267T+H272V+F276A+S279G+Y281P+K308Q+K309E+K312QandQ3F+H15Y+A49W+N59E+V140I+S163P+S186R+Q215K+S225G+N227T+E232D+G235W+N252P+N260Q+N267T+H272V+S279D+Y281P+K308Q+K309E+K312Q,wherein the variant has at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 95%, suchas at least 96%, at least 97%, at least 98%, or at least 99%, but lessthan 100% sequence identity to the polypeptide shown in SEQ ID NO: 1,wherein each position corresponds to the position of the polypeptideshown in SEQ ID NO: 1 (numbering according to SEQ ID NO: 1).

In one preferred embodiment of the invention the dispersin variant ofthe invention comprises one of the substitution sets selected from thegroup consisting of:

A49W+F276A+K308E;

A49W+N227T+A269C+Y282C+K308E;

A49W+N227T+F276A;

A49W+N227T+F276A+K308E;

A49W+S186R+N227T+F276A;

A49W+S186R+N227T+F276A+K308E;

A49W+S186R+N227T+K308E;

A49W+Y124R+N227T+F276A+K308E;

A49W+Y124R+S186R+N227T+F276A;

A49W+Y124R+S186R+N227T+F276A+K308E;

A49W+Y124R+S186R+N227T+K308E;

S163P+N227T+F276A+K308E;

S163P+Q215K+N227T+N252P+N267T+F276A;

S163P+Q215K+N227T+N252P+N267T+F276A+K308E;

S163P+Q215K+N227T+N252P+N267T+F276A+K309E;

S163P+Q215K+N227T+N252P+N267T+F276A+K312E;

S163P+Q215K+N252P;

S186R+F276A+K308E;

S186R+N227T+F276A;

S186R+N227T+K308E;

Y124R+S186R+N227T+A269C+F276A+Y282C+K308E; andY124R+S186R+N227T+F276A+K308E, wherein the variant has at least 60%, atleast 65%, at least 70%, at least 75%, at least 80%, at least 85%, atleast 90%, at least 95%, such as at least 96%, at least 97%, at least98%, or at least 99%, but less than 100% sequence identity to thepolypeptide shown in SEQ ID NO: 1, wherein each position corresponds tothe position of the polypeptide shown in SEQ ID NO: 1 (numberingaccording to SEQ ID NO: 1).

The dispersin variants above preferably has at least 60%, at least 65%,at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98%, or at least99%, but less than 100% sequence identity to the polypeptide shown inSEQ ID NO: 1.

In one preferred embodiment the dispersin comprises the polypeptideshown in SEQ ID NO: 17 or a polypeptide having at least at least 60%, atleast 65%, at least 70%, at least 75%, at least 80%, at least 85%, atleast 90%, at least 95%, such as at least 96%, at least 97%, at least98%, or at least 99%, but less than 100% sequence identity hereto.

In some aspects, the dispersin variant comprises one or moresubstitution sets selected from the group consisting of: Q3F+A49W,Q3F+N59E , Q3F+S163P, Q3F+S186R, Q3F+Q215K, Q3F+S225G, Q3F+N227T,Q3F+N252P, Q3F+N267T, Q3F+F276A, Q3F+Y281P, Q3F+K308E, Q3F+K309E,Q3F+K312E, A49W+N59E, A49W+S163P, A49W+S186R, A49W+Q215K, A49W+S225G,A49W+N227T, A49W+N252P, A49W+N267T, A49W+F276A, A49W+Y281P, A49W+K308E,A49W+K309E, A49W+K312E, N59E+S163P, N59E+S186R, N59E+Q215K, N59E+S225G,N59E+N227T, N59E+N252P, N59E+N267T, N59E+F276A, N59E+Y281P, N59E+K308E,N59E+K309E, N59E+K312E, S163P+S186R, S163P+Q215K, S163P+S225G,S163P+N227T, S163P+N252P, S163P+N267T, S163P+F276A, S163P+Y281P,S163P+K308E, S163P+K309E, S163P+K312E, S186R+Q215K, S186R+S225G,S186R+N227T, S186R+N252P, S186R+N267T, S186R+F276A, S186R+Y281P,S186R+K308E, S186R+K309E, S186R+K312E, Q215K+S225G, Q215K+N227T,Q215K+N252P, Q215K+N267T, Q215K+F276A, Q215K+Y281P, Q215K+K308E,Q215K+K309E, Q215K+K312E, S225G+N227T, S225G+N252P, S225G+N267T,S225G+F276A, S225G+Y281P, S225G+K308E, S225G+K309E, S225G+K312E,N227T+N252P, N227T+N267T, N227T+F276A, N227T+Y281P, N227T+K308E,N227T+K309E, N227T+K312E, N252P+N267T, N252P+F276A, N252P+Y281P,N252P+K308E, N252P+K309E, N252P+K312E, N267T+F276A, N267T+Y281P,N267T+K308E, N267T+K309E, N267T+K312E, F276A+Y281P, F276A+K308E,F276A+K309E, F276A+K312E, Y281P+K308E, Y281P+K309E, Y281P+K312E,K308E+K309E, K308E+K312E and K309E+K312E, wherein each substitutionprovides a dispersin variant having an increase in stability measured ashalf-life improvement factor, HIF or residual activity ratio, RAR, of atleast 1.05, such as 1.08, such as 1.1, such as 1.15, such as 1.2, suchas 1.25, such as 1.3, such as 1.4, such as 1.5, such as 1.6, such as1.7, such as 1.8, such as 1.9, such as 2, such as 3, such as 4, such as5 or such as at least 10 compared to the parent dispersin e.g. adispersin comprising the polypeptide having the amino acid sequenceshown in SEQ ID NO: 1, wherein the dispersin variant has at least 60%,at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, atleast 90%, at least 95%, such as at least 96%, at least 97%, at least98%, or at least 99%, but less than 100% sequence identity to thepolypeptide shown in SEQ ID NO: 1, wherein each position corresponds tothe position of the polypeptide shown in SEQ ID NO: 1 (numberingaccording to SEQ ID NO: 1).

In one particular embodiment, the dispersin variant of the inventioncomprises at least one di-sulfide bridge. In one embodiment, thedispersin variant of the invention comprises at least one of thedisulfide bridge: A269C-Y282C.

In one embodiment the dispersin variant comprises any of the followingsubstitutions and disulfide bridges:

A49W+A269C+F276A+Y282C+K308E,

A49W+N227T+A269C+F276A+Y282C,

A49W+N227T+A269C+F276A+Y282C+K308E,

A49W+S186R+N227T+A269C+F276A+Y282C,

A49W+S186R+N227T+A269C+F276A+Y282C+K308E,

A49W+S186R+N227T+A269C+Y282C+K308E,

A49W+Y124R+N227T+A269C+F276A+Y282C+K308E,

A49W+Y124R+S186R+N227T+A269C+F276A+Y282C,

A49W+Y124R+S186R+N227T+A269C+F276A+Y282C+K308E,

A49W+Y124R+S186R+N227T+A269C+Y282C+K308E,

N227T+A269C+F276A+Y282C+K308E,

S186R+A269C+F276A+Y282C+K308E,

S186R+N227T+A269C+F276A+Y282C,

S186R+N227T+A269C+F276A+Y282C+K308E,

S186R+N227T+A269C+Y282C+K308E or

Y124R+S186R+N227T+A269C+F276A+Y282C+K308E.

Preferably the dispersin variants have at least 60%, at least 65%, atleast 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 95%, such as at least 96%, at least 97%, at least 98% or 100%sequence identity to SEQ ID NO: 1.

The dispersin variant may also comprise any of the followingalterations: D2E, D2S, D2T, Q3A, Q3*, T17A, E19T, E19H, E19W, E19L,K22R, K22N, K22M, K22S, D26*, S29G, Y30C, Y30S, Y30E, Y30G, N31W, N31G,G32N, N34*, N34M, N43D, N43F, E44N, E44S, E44I, S50C, E51D, Y52F, G54D,G54R, S56D, S56A, S57N, S57L, E58R, E58K, E58W, E58P, N59Q, N59K, N59I,N59G, T60I, T60M, T6OR, T60A, N61E, N62K, N62M, N62V, N62G, T63Y, T63I,T63K, K67W, N68M, N68A, L71K, L71Q, L71E, L71D, S72P, A75R, A75W, A75G,A75N, A75M, D79R, D79L, D79N, D79E, D79W, K801, K80D, K80Y, D81M, D81I,D81*, D81E, K95M, G96A, E99V, E99M, K102A, K102Q, K103D, K103C, K103P,K103T, K103N, K103I, K103S, K104A, K104V, K104*, D105*, V1061, V106P,V106*, V106S, V106F, K107R, K107G, K107N, K107H, K107Q, L108M, N110W,D111L, D111K, D111H, V113*, V113R, D115V, Y116*, Y116E, S117W, S117V,S117R, E118C, E118F, E118H, E119*, E119T, E119Y, E119M, E119H, Y124A,Y124S, Y124K, Y124I, Y124F, D125G, R127V, R127G, V128W, V128L, D131R,D131*, N134R, N134M, Q135H, Q135W, D138A, D138C, D138I, D138E, D138K,D142M, D142L, D142T, D142V, D142*, Y145A, Y145K, Y145R, Y145G, P1471,P147*, P147N, P147E, K148M, K148R, K148G, K148P, K148N, F149Y, F149E,E150I, G151M, G151R, G151V, G151T, G151E, G151F, K152V, K152W, K152S,G164F, G164Q, G164L, G164W, G164R, G164A, G164C, V167N, H168D, H168M,H168A, H168W, H168S, L170I, D171F, D171N, D171M, D174L, D174T, D174A,D174R, N177D, N177*, Q178E, Q178H, S181H, S181M, S181R, S181G, S181F,K184P, K184M, K184D, E185R, E185D, E185N, E185M, S186A, S186H, S186P,K187N, K187F, K187Y, E189M, S1991, S199E, S199N, S199A, S199R, E200Q,E200H, E200I, E200Y, A203N, A203I, N204A, N204Y, D207M, D207L, D207P,D207H, S208M, S208N, S208P, S208R, S217L, S217A, S217R, T218E, T218C,T218K, G222N, G222K, E224G, E224L, D230M, N233I, W234K, W234N, W234F,G235D, S237A, S237T, S237P, Y244V, L249D, L249I, L249T, S251Q, S251K,S251D, S251T, N252T, N252W, N252Y, G253E, G253*, G253R, G253I, G253A,F254T, F254K, F254N, F254V, F254A, F254*, T255E, Q256V, E257D, N260D,N260H, E261N, Q262L, Q262W, Q262H, N267V, W268Q, W268V, A271F, H272*,H272N, H272T, H272S, I278T, D280M, Y282P, Y282D, H283K, A284L, E286D,S288F, S288W, S288A, S288Q, S288E, E300N, E300L, H301L, H301V, H301M,T303D, T303M, T303E, T303N, Q307L, K308A, K308D, K309V, L311M, R319A orR319L, preferably wherein the dispersin variants have at least 60%, atleast 65%, at least 70%, at least 75%, at least 80%, at least 85%, atleast 90%, at least 95%, such as at least 96%, at least 97%, at least98% or 100% sequence identity to SEQ ID NO: 1.

The dispersin variant may also comprise any of the followingalterations: N31W N31G, S50C, N59I, N59Q, G96A, Y124A, V128L, D138A,F149Y, S181H, K184P, K184M, E189M, N204A, D207N, S208A, Q262L, Q262W,H283K, E300N, T303G, T303C or K309V, preferably the dispersin variantshave at least 60%, at least 65%, at least 70%, at least 75%, at least80%, at least 85%, at least 90%, at least 95%, such as at least 96%, atleast 97%, at least 98% or 100% sequence identity to SEQ ID NO: 1.

Examples of conservative substitutions are within the groups of basicamino acids (arginine, lysine and histidine), acidic amino acids(glutamic acid and aspartic acid), polar amino acids (glutamine andasparagine), hydrophobic amino acids (leucine, isoleucine and valine),aromatic amino acids (phenylalanine, tryptophan and tyrosine), and smallamino acids (glycine, alanine, serine, threonine and methionine). Aminoacid substitutions that do not generally alter specific activity areknown in the art and are described, for example, by H. Neurath and R.L.Hill, 1979, In, The Proteins, Academic Press, New York. Commonsubstitutions are Ala/Ser, Val/Ile, Asp/Glu, Thr/Ser, Ala/Gly, Ala/Thr,Ser/Asn, Ala/Val, Ser/Gly, Tyr/Phe, Ala/Pro, Lys/Arg, Asp/Asn, Leu/Ile,Leu/Val, Ala/Glu, and Asp/Gly.

Essential amino acids in a polypeptide can be identified according toprocedures known in the art, such as site-directed mutagenesis oralanine-scanning mutagenesis (Cunningham and Wells, 1989, Science 244:1081-1085). In the latter technique, single alanine mutations areintroduced at every residue in the molecule, and the resultant mutantmolecules are tested for dispersin activity to identify amino acidresidues that are critical to the activity of the molecule. See also,Hilton et al., 1996, J. Biol. Chem. 271: 4699-4708. The active site ofthe enzyme or other biological interaction can also be determined byphysical analysis of structure, as determined by such techniques asnuclear magnetic resonance, crystallography, electron diffraction, orphotoaffinity labelling, in conjunction with mutation of putativecontact site amino acids. See, for example, de Vos et al., 1992, Science255: 306-312; Smith et al., 1992, J. Mol. Biol. 224: 899-904; Wlodaveret al., 1992, FEBS Lett. 309: 59-64.

Parent Dispersin

Preferably, the dispersin parent is obtained from a microorganism andthe dispersin is a microbial enzyme. The dispersin is preferably offungal or bacterial origin. The dispersin parent is preferablyobtainable from Terribacillus e.g. Terribacillus saccharophilus,Terribacillus goriensis or Terribacillus saccharophilus.

The parent dispersin is preferably selected from the group consistingof:

-   -   a) a polypeptide comprising the amino acid sequence shown in SEQ        ID NO: 1 or is a polypeptide having at least 60%, at least 65%,        at least 70%, at least 75%, at least 80%, at least 85%, at least        90%, at least 95%, such as at least 96%, at least 97%, at least        98% or 100% sequence identity hereto,    -   b) a polypeptide comprising the amino acid sequence shown in SEQ        ID NO:2 or is a polypeptide having at least 60%, at least 65%,        at least 70%, at least 75%, at least 80%, at least 85%, at least        90%, at least 95%, such as at least 96%, at least 97%, at least        98% or 100% sequence identity hereto,    -   c) a polypeptide comprising the amino acid sequence shown in SEQ        ID NO:3 or is a polypeptide having at least 60%, at least 65%,        at least 70%, at least 75%, at least 80%, at least 85%, at least        90%, at least 95%, such as at least 96%, at least 97%, at least        98% or 100% sequence identity hereto,    -   d) a polypeptide comprising the amino acid sequence shown in SEQ        ID NO:4 or is a polypeptide having at least 60%, at least 65%,        at least 70%, at least 75%, at least 80%, at least 85%, at least        90%, at least 95%, such as at least 96%, at least 97%, at least        98% or 100% sequence identity hereto, and    -   e) a polypeptide comprising the amino acid sequence shown in SEQ        ID NO:5 or is a polypeptide having at least 60%, at least 65%,        at least 70%, at least 75%, at least 80%, at least 85%, at least        90%, at least 95%, such as at least 96%, at least 97%, at least        98% or 100% sequence identity hereto.        The parent dispersins preferably belong to the Terribacillus        clade and comprises one or more of the conserved motifs: GXDE        (SEQ ID NO:8), [EQ][NRSHA][YVFL][AGSTC][IVLF][EAQYN][SN] (SEQ ID        NO:9),[VIM][LIV]G[GAV]DE[VI][PSA] SEQ ID NO: 10),        [WND[SQR][IVL][TLVM] (SEQ ID NO: 11), QSTL (SEQ ID NO: 12),        NKFFY (SEQ ID NO: 13) or NLD[DR]S (SEQ ID NO: 14),        An overview of the Terribacillus clade is provided in the table        below. The Terribacillus clade comprises homologous sequences.        The polypeptides with hexosaminidase activity of the present        invention having the mature amino acid sequences SEQ ID Nos 1-5        can be pairwise aligned using the Needleman-Wunsch algorithm        (Needleman and Wunsch, 1970, J. Mol. Biol. 48: 443-453). The        percent identities resulting from such alignments are shown        below.

SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID NO: 5 NO: 4 NO: 3 NO: 2 NO: 1 10075.9 81.8 82.4 81.2 SEQ ID NO: 5 75.9 100 77.5 76.5 77.5 SEQ ID NO: 481.8 77.5 100 95.1 95.7 SEQ ID NO: 3 82.4 76.5 95.1 100 93.5 SEQ ID NO:2 81.2 77.5 95.7 93.5 100 SEQ ID NO: 1The table shows that the polypeptides of the invention share closesequence relatedness. The polypeptides comprising the amino acidssequences of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4 and SEQID NO:5 belongs to a subclade of the Dispersin clade. These polypeptidesshare more than 90% pairwise sequence identity and are close related toeach other.The polypeptides of the invention all lie within the same clade, theTerribacillus clade, and all have common functional features includingcleaning properties in the presence of detergents. In one aspect theparent dispersins belongs to the Terribacillus clade and comprises oneor more of the motif(s) GXDE (SEQ ID NO:8),[EQ][NRSHA][YVFL][AGSTC][IVLF][EAQYN][SN] (SEQ ID NO:9),[VIM][LIV]G[GAV]DE[VI][PSA] (SEQ ID NO: 10), WND[SQR][IVL][TLVM] (SEQ IDNO: 11), QSTL (SEQ ID NO: 12), NKFFY (SEQ ID NO: 13) or NLD[DR]S (SEQ IDNO: 14).

Use and Methods

The dispersin variants of the present invention may be added to and thusbecome a component of a cleaning e.g. detergent composition.

The cleaning composition of the present invention may be formulated, forexample, as a hand or machine laundry detergent composition including alaundry additive composition suitable for pre-treatment of stainedfabrics and a rinse added fabric softener composition, or be formulatedas a detergent composition for use in general household hard surfacecleaning operations, or be formulated for hand or machine dishwashingoperations.

In a specific aspect, the present invention provides a detergentadditive comprising a dispersin variant of the present invention asdescribed herein.

The cleaning compositions may be used for cleaning an item such as atextile or a hard surface in e.g. laundry and dish wash processes.

One aspect relates to a method of cleaning an item comprising a) addinga dispersin variant according to invention to a cleaning composition andb) washing item with the composition, wherein the item is a textile or ahard surface.

One aspect relates to a method of cleaning an item comprising;

-   -   a) adding a dispersin variant, comprising an alteration,        preferably substitution at one or more positions corresponding        to positions 2, 3, 12, 15, 17, 18, 19, 22, 23, 24, 25, 26, 30,        32, 34, 43, 44, 45, 49, 52, 54, 56, 57, 58, 59, 60, 62, 63, 67,        68, 71, 72, 74, 77, 79, 80, 81, 82, 90, 99, 100, 103, 104, 105,        106, 107, 110, 111, 113, 114, 116, 117, 118, 119, 120, 122, 123,        124, 125, 126, 127, 128, 131, 135, 138, 139, 140, 142, 145, 147,        148, 149, 150, 151, 152, 163, 164, 167, 168, 170, 171, 173, 174,        175, 177, 178, 179, 181, 185, 186, 187, 188, 189, 199, 200, 203,        204, 205, 207, 208, 210, 215, 217, 218, 221, 222, 224, 225, 227,        230, 232, 233, 234, 235, 237, 244, 249, 251, 252, 253, 254, 256,        260, 261, 262, 263, 264, 265, 267, 268, 270, 271, 272, 273, 274,        276, 278, 279, 280, 281, 282, 283, 284, 287, 288, 290, 291, 296,        300, 301, 303, 304, 305, 306, 308, 309, 312, 314, 315, 319, 321        or 323 of the polypeptide of SEQ ID NO: 1, wherein the dispersin        variant has beta-1,6 N-acetylglucosaminidase activity, wherein        the dispersin variant has at least 80%, at least 85%, at least        90%, at least 95%, at least 96%, at least 97%, at least 98%, or        at least 99%, but less than 100% sequence identity to the        polypeptide shown in SEQ ID NO: 1, to a cleaning composition and    -   b) washing item with the composition, wherein the item is a        textile or a hard surface.

One aspect relates to a method of cleaning an item comprising;

-   -   a) adding a dispersin variant, comprising an alteration,        preferably substitution at one or more positions corresponding        to positions 31, 33, 36, 73, 75, 94, 141, 144, 241, 277, 294,        297 or 299 of the polypeptide of SEQ ID NO: 1, wherein the        dispersin variant has beta-1,6 N-acetylglucosaminidase activity,        wherein the dispersin variant has at least 80%, at least 85%, at        least 90%, at least 95%, at least 96%, at least 97%, at least        98%, or at least 99%, but less than 100% sequence identity to        the polypeptide shown in SEQ ID NO: 1, to a cleaning composition        and    -   b) washing item with the composition, wherein the item is a        textile or a hard surface.

One aspect relates to a method of cleaning an item comprising a) addinga dispersin variant, wherein the variant comprises any of thealterations selected from the group consisting of: D2A, D2L, D2N, D2R,D2V, D2W, Q3F, Q3I, Q3L, Q3M, Q3P, Q3V, Q3Y, Q3T, S12A, H15F, H15Y,T17W, T17C, T17E, T17F, T17M, T17R, T17V, V18L, E19D, E19K, E19N, E19P,K22A, K22M, K22V, S23A, S23C, S23E, S23I, S23L, S23R, S23T, S23V, L24I,V25R, D26M, Y30*, Y30A, Y30D, Y30L, Y30M, Y30N, Y30R, Y30T, Y30V, G32L,G32M, G32R, N34D, N43*, N43H, N43L, E44*, N45D, N45L, N45V, A49W, A49Y,Y52*, Y52M, G54L, G54M, G54N, S56T, S56W, S57W, E58N, N59A, N59C, N59D,N59E, N59F, N59M, N59R, N59V, N59W, T60V, N62C, N62D, N62H, N62Q, N62W,T63C, T63D, T63L, T63N, T63R, T63V, K67A, K67L, N68L, N68Q, L71H, L71N,L71R, L71V, L71W, S72*, S72C, S72D, S72E, S72F, S72G, S72I, S72M, S72N,S72R, S72T, S72Y, I74L, S77A, D79V, K80*, K80E, K80H, K80L, K80N, K80Q,K80R, K80V, K80W, D81A, D81G, D81L, D81N, D81R, D81S, D81T, D81V, D81W,I82V, L90F, E99Q, E99R, L100S, K103A, K103R, K103V, K104N, K104W, D105N,V106A, V106D, V106E, V106H, V106K, V106L, V106M, V106N, V106Q, V106R,V106W, V106Y, K107A, K107C, K107L, K107M, K107T, K107V, K107W, N110M,N110R, N110V, D111A, D111E, D111M, D111N, D111Q, D111R, D111V, D111W,V113T, T114C, T114S, Y116D, Y116N, Y116R, S117*, S117D, S117H, S117N,S117P, E118*, E118A, E118D, E118G, E118L, E119G, E119W, T120I, T120L,T120M, T120V, T120W, D122*, D122H, D122R, Y123W, Y124C, Y124H, Y124I,Y124K, Y124L, Y124M, Y124N, Y124Q, Y124R, Y124T, Y124V, Y124W, D125C,D125G, D125H, D125K, D125Q, D125R, N126V, R127D, R127H, R127K, R127L,R127M, R127Q, R127W, V128A, V128C, V128D, V128L, V128T, D131V, Q135*,Q135A, Q135D, Q135E, Q135K, Q135M, Q135Y, D138K, D138L, D138M, D138N,D138Q, D138R, D138S, D138V, D138W, E139W, V140I, D142R, D142W, Y145*,Y145H, Y145L, Y145N, Y145V, P147A, P147C, P147D, P147F, P147G, P147L,P147M, P147R, P147S, P147T, P147V, K148A, K148D, K148L, K148V, F149L,F149M, F149N, E150A, E150D, E150H, E150K, E150L, E150M, E150N, E150R,E150V, E150W, E150Y, G151A, G151C, G151D, G151L, G151N, G151P, G151S,G151W, K152D, K152L, K152R, S163P, G164D, G164E, G164H, G164S, G164V,V167A, V167D, V167E, V167L, V167P, V167Q, V167R, V167W, H168N, L170A,L170D, L170E, L170F, L170H, L170K, L170M, L170N, L170P, L170Q, L170R,L170S, L170V, L170W, L170Y, D171A, D171C, D171E, D171K, D171L, D171M,D171Q, D171R, D171V, D171W, D171Y, I173C, D174H, D174M, D174N, D174V,D174W, F175Y, N177M, Q178*, Q178A, Q178K, Q178R, Q178W, I179T, S181C,S181D, S181F, S181G, S181K, S181N, S181P, S181Q, S181T, S181V, S181W,E185A, E185M, E185R, E185V, E185W, S186D, S186E, S186H, S186I, S186K,S186L, S186M, S186N, S186Q, S186R, S186V, S186W, K187C, K187D, K187G,K187R, K187S, K187V, K187W, Y188P, E189L, E189V, E189W, S199C, S199L,S199M, S199Y, E200D, E200F, E200K, E200L, E200M, E200N, E200R, E200W,A203C, A203D, A203E, A203G, A203L, A203M, A203P, A203Q, A203R, A203S,A203T, A203V, A203W, N204L, N204M, N204V, N204W, N204Y, L205I, D207A,D207C, D207E, D207G, D207K, D207N, D207Q, D207R, D207S, D207V, D207W,S208A, S208C, S208D, S208G, S208L, S208Q, S208T, S208V, S208W, S210T,Q215K, Q215R, Q215M, Q215L, Q215*, S217V, T218A, T218L, T218Q, T218R,T218V, S221N, G222D, E224A, E224P, S225G, N227A, N227Q, N227R, N227S,N227T, N227K, D230*, D230N, D230R, D230T, D230W, E232D, E232V, N233D,N233E, N233H, N233Q, N233R, N233W, W234R, G235W, G235A, G235E, G235F,G235H, G235I, G235L, G235M, G235N, G235P, G235S, G235V, S237C, S237G,S237M, S237N, S237W, S237Y, Y244C, Y244E, Y244M, L249H, L249K, L249Q,L249R, L249W, L249Y, S251A, S251L, S251N, S251R, S251W, N252P, N252C,G253D, G253W, F254I, F254L, F254M, F254Y, Q256D, Q256E, Q256R, N260*,N260A, N260C, N260E, N260I, N260K, N260L, N260M, N260Q, N260R, N260T,N260V, N260W, N260Y, E261*, E261A, E261D, E261R, E261W, Q262*, Q262F,Q262H, Q262W, Q262Y, M263K, M263L, M263Q, D264*, D264C, D264E, D264N,Y265F, N267S, N267T, W268C, W268E, W268M, W268R, Y270F, A271D, A271G,H272D, H272I, H272M, H272P, H272V, H272W, N273W, K274R, K274A, K274H,F276A, F276C, F276K, F276N, F276G, F276L, F276M, F276P, F276S, F276V,F276W, I278A, I278K, I278N, I278Q, I278V, S279C, S279D, S279E, S279G,S279N, D280C, D280E, Y281*, Y281A, Y281C, Y281H, Y281K, Y281N, Y281P,Y281R, Y282E, Y282N, H283I, A284I, A284L, A284N, A284P, A284T, A284V,T287N, S288P, S288D, S288K, S288N, V290I, K291L, K291R, K291V, T296C,E300A, E300D, H301C, H301N, H301R, T303A, T303C, T303G, T303K, T303Q,T303R, T303W, D304C, D304M, L305M, L305N, S306C, K308A, K308D, K308E,K308G, K308I, K308L, K308Q, K308S, K308T, K308V, K308Y, K309A, K309C,K309D, K309E, K309G, K309H, K309L, K309M, K309N, K309Q, K309S, K309T,K309I, K312A, K312E, K312L, K312M, K312N, K312Q, K312S, K312W, E314I,E314L, E314V, L315I, L315V, R319A, Y321F and N323R compared to thedispersin shown in SEQ ID NO: 1 to a cleaning composition and b) washingitem with the composition, wherein the item is a textile or a hardsurface.

The dispersin variants of the invention are suitable for use in cleaningsuch as laundry. Thus, some aspect of the invention relates to a methodfor laundering an item, wherein the method comprises the steps of:

-   -   a. Exposing an item to a wash liquor comprising a dispersin        variant of the invention;    -   b. Completing at least one wash cycle; and    -   c. Optionally rinsing the item, wherein the item is a textile or        a hard surface.

The pH of the liquid solution is in the range of 1 to 11, such as in therange 5.5 to 11, such as in the range of 7 to 9, in the range of 7 to 8or in the range of 7 to 8.5.

The wash liquor may have a temperature in the range of 5° C. to 95° C.,or in the range of 10° C. to 80° C., in the range of 10° C. to 70° C.,in the range of 10° C. to 60° C., in the range of 10° C. to 50° C., inthe range of 15° C. to 40° C. or in the range of 20° C. to 30° C. Insome aspects the temperature of the wash liquor is 30° C.

The dispersin variants of the invention may be added to a wash liquor.

The concentration of the dispersin variant enzyme in the wash liquor istypically in the range of 0.00001-1000 ppm enzyme protein, such as inthe range of 0.00004-100 ppm enzyme protein, such as in the range of0.00008-100 enzyme protein, in the range of 0.0001-100 enzyme protein,in the range of 0.0002-100 enzyme protein, in the range of 0.0004-100enzyme protein, in the range of 0.0008-100 enzyme protein, in the rangeof 0.001-100 ppm enzyme protein, in the range of 0.01-100 ppm enzymeprotein, in the range of 1-1000 ppm enzyme protein, preferably in therange of 0.05-50 ppm enzyme protein, more preferably in the range of0.1-50 ppm enzyme protein, more preferably in the range of 0.1-30 ppmenzyme protein, more preferably in the range of 0.5-20 ppm enzymeprotein, and most preferably in the range of 0.5-10 ppm enzyme protein.

In some aspects the dispersin variants of the present invention areeffective in preventing and/or reducing the malodor. The presence ofcomplex organic stains such as body soil e.g. cell debris, sebum or EPSattach to e.g. laundry items which become sticky and therefore soiladheres to the sticky areas. This soil s to the laundry has showndifficult to reduce by commercially available detergent compositions.Further, when dirty laundry items are washed together with less dirtylaundry items the dirt present in the wash liquor tend to adhere to thelaundry (e.g. by re-deposition) in particular if the laundry is stickyas described above. As a result, hereof, the laundry item is more“soiled” after wash than before wash. In some aspects, the dispersinvariants of the invention have improved cleaning properties compared tothe parent dispersin and in some aspects, the dispersin variants of theinvention reduce stickiness and/or re-deposition.

In some aspects, the invention relates to the use of a dispersin variantaccording to the invention for cleaning of an item, wherein the item isa fabric or a hard surface.

Further, the invention relates to the use of a dispersin variantaccording to the invention for preventing and/or reducing the adherenceof soil to an item. In some aspect, the item is textile. When the soildoes not adhere to the item, the item appears cleaner. Thus, theinvention further relates to the use of a dispersin variant according tothe invention for maintaining or improving the whiteness of the item.

The present invention further relates to detergent compositionscomprising a dispersin variant according to the invention preferablywith a detergent adjunct ingredient. The detergent compositioncomprising a dispersin variant according to the invention may be usedfor deep cleaning of an item, for preventing and/or reducing thestickiness of an item, for pretreating stains on the item, forpreventing and/or reducing redeposition of soil during a wash cycle, forpreventing and/or reducing adherence of soil to an item, for maintainingor improving the whiteness of an item and/or for preventing and/orreducing malodor from an item.

Preparation of Variants

The present invention also relates to a method for obtaining a dispersinvariant having at least one improved property compared to the parentdispersin e.g. compared to the polypeptide shown in SEQ ID NO: 1.

One aspect of the invention relates to method for obtaining a dispersinvariant, comprising;

-   -   a) introducing into a parent dispersin one or more alterations        at a position selected from the group consisting of: 2, 3, 12,        15, 17, 18, 19, 22, 23, 24, 25, 26, 30, 32, 34, 43, 44, 45, 49,        52, 54, 56, 57, 58, 59, 60, 62, 63, 67, 68, 71, 72, 74, 77, 79,        80, 81, 82, 90, 99, 100, 103, 104, 105, 106, 107, 110, 111, 113,        114, 116, 117, 118, 119, 120, 122, 123, 124, 125, 126, 127, 128,        131, 135, 138, 139, 140, 142, 145, 147, 148, 149, 150, 151, 152,        163, 164, 167, 168, 170, 171, 173, 174, 175, 177, 178, 179, 181,        185, 186, 187, 188, 189, 199, 200, 203, 204, 205, 207, 208, 210,        215, 217, 218, 221, 222, 224, 225, 227, 230, 233, 234, 235, 237,        244, 249, 251, 252, 253, 254, 256, 260, 261, 262, 263, 264, 265,        267, 268, 270, 271, 272, 273, 274, 276, 278, 279, 280, 281, 282,        283, 284, 287, 288, 290, 291, 296, 300, 301, 303, 304, 305, 306,        308, 309, 312, 314, 315, 319, 321 and 323; and    -   b) recovering the variant, wherein the variant has        hexosaminidase, preferably beta-1,6 N-acetylglucosaminidase        activity.

One aspect of the invention relates to method for obtaining a dispersinvariant, comprising;

-   -   a) introducing into a parent dispersin one or more alterations        at a position selected from the group consisting of: 2, 3, 12,        15, 17, 18, 22, 23, 24, 25, 30, 49, 56, 57, 59, 62, 63, 68, 72,        74, 77, 82, 90, 99, 100, 106, 114, 123, 124, 125, 135, 138, 140,        163, 167, 170, 171, 173, 174, 175, 178, 179, 181, 185, 186, 187,        188, 189, 199, 203, 204, 205, 207, 210, 215, 218, 221, 225, 227,        235, 237, 244, 252, 256, 260, 262, 263, 264, 265, 267, 270, 272,        273, 274, 276, 278, 279, 280, 281, 282, 283, 284, 288, 290, 291,        296, 303, 304, 305, 306, 308, 309, 312, 314, 315, 319, 321, 322        and 323; and    -   b) recovering the variant, wherein the variant has        hexosaminidase, preferably beta-1,6 N-acetylglucosaminidase        activity.

One aspect of the invention relates to method for obtaining a dispersinvariant, comprising;

-   -   a) introducing into a parent dispersin one or more alterations        at a position selected from the group consisting of: 31, 33, 36,        73, 75, 94, 141, 144, 241, 277, 294, 297, 299 and    -   b) recovering the variant, wherein the variant has        hexosaminidase, preferably beta-1,6 N-acetylglucosaminidase        activity.

One aspect of the invention relates to method for obtaining a dispersinvariant, comprising;

-   -   a) introducing into a parent dispersin one or more alterations        selected from the group consisting of: D2A, D2L, D2N, D2R, D2V,        D2W, Q3F, Q3I, Q3L, Q3M, Q3P, Q3V, Q3Y, Q3T, S12A, H15F, H15Y,        T17W, T17C, T17E, T17F, T17M, T17R, T17V, V18L, E19D, E19K,        E19N, E19P, K22A, K22M, K22V, S23A, S23C, S23E, S23I, S23L,        S23R, S23T, S23V, L24I, V25R, D26M, Y30*, Y30A, Y30D, Y30L,        Y30M, Y30N, Y30R, Y30T, Y30V, G32L, G32M, G32R, N34D, N43*,        N43H, N43L, E44*, N45D, N45L, N45V, A49W, A49Y, Y52*, Y52M,        G54L, G54M, G54N, S56T, S56W, S57W, E58N, N59A, N59C, N59D,        N59E, N59F, N59M, N59R, N59V, N59W, T60V, N62C, N62D, N62H,        N62Q, N62W, T63C, T63D, T63L, T63N, T63R, T63V, K67A, K67L,        N68L, N68Q, L71H, L71N, L71R, L71V, L71W, S72*, S72C, S72D,        S72E, S72F, S72G, S72I, S72M, S72N, S72R, S72T, S72Y, I74L,        S77A, D79V, K80*, K80E, K80H, K80L, K80N, K80Q, K80R, K80V,        K80W, D81A, D81G, D81L, D81N, D81R, D81S, D81T, D81V, D81W,        I82V, L90F, E99Q, E99R, L100S, K103A, K103R, K103V, K104N,        K104W, D105N, V106A, V106D, V106E, V106H, V106K, V106L, V106M,        V106N, V106Q, V106R, V106W, V106Y, K107A, K107C, K107L, K107M,        K107T, K107V, K107W, N110M, N110R, N110V, D111A, D111E, D111M,        D111N, D111Q, D111R, D111V, D111W, V113T, T114C, T114S, Y116D,        Y116N, Y116R, S117*, S117D, S117H, S117N, S117P, E118*, E118A,        E118D, E118G, E118L, E119G, E119W, T120I, T120L, T120M, T120V,        T120W, D122*, D122H, D122R, Y123W, Y124C, Y124H, Y124I, Y124K,        Y124L, Y124M, Y124N, Y124Q, Y124R, Y124T, Y124V, Y124W, D125C,        D125G, D125H, D125K, D125Q, D125R, N126V, R127D, R127H, R127K,        R127L, R127M, R127Q, R127W, V128A, V128C, V128D, V128L, V128T,        D131V, Q135*, Q135A, Q135D, Q135E, Q135K, Q135M, Q135Y, D138K,        D138L, D138M, D138N, D138Q, D138R, D138S, D138V, D138W, E139W,        V140I, D142R, D142W, Y145*, Y145H, Y145L, Y145N, Y145V, P147A,        P147C, P147D, P147F, P147G, P147L, P147M, P147R, P147S, P147T,        P147V, K148A, K148D, K148L, K148V, F149L, F149M, F149N, E150A,        E150D, E150H, E150K, E150L, E150M, E150N, E150R, E150V, E150W,        E150Y, G151A, G151C, G151D, G151L, G151N, G151P, G151S, G151W,        K152D, K152L, K152R, S163P, G164D, G164E, G164H, G164S, G164V,        V167A, V167D, V167E, V167L, V167P, V167Q, V167R, V167W, H168N,        L170A, L170D, L170E, L170F, L170H, L170K, L170M, L170N, L170P,        L170Q, L170R, L170S, L170V, L170W, L170Y, D171A, D171C, D171E,        D171K, D171L, D171M, D171Q, D171R, D171V, D171W, D171Y, I173C,        D174H, D174M, D174N, D174V, D174W, F175Y, N177M, Q178*, Q178A,        Q178K, Q178R, Q178W, I179T, S181C, S181D, S181F, S181G, S181K,        S181N, S181P, S181Q, S181T, S181V, S181W, E185A, E185M, E185R,        E185V, E185W, S186D, S186E, S186H, S186I, S186K, S186L, S186M,        S186N, S186Q, S186R, S186V, S186W, K187C, K187D, K187G, K187R,        K187S, K187V, K187W, Y188P, E189L, E189V, E189W, S199C, S199L,        S199M, S199Y, E200D, E200F, E200K, E200L, E200M, E200N, E200R,        E200W, A203C, A203D, A203E, A203G, A203L, A203M, A203P, A203Q,        A203R, A203S, A203T, A203V, A203W, N204L, N204M, N204V, N204W,        N204Y, L205I, D207A, D207C, D207E, D207G, D207K, D207N, D207Q,        D207R, D207S, D207V, D207W, S208A, S208C, S208D, S208G, S208L,        S208Q, S208T, S208V, S208W, S210T, Q215K, Q215R, Q215M, Q215L,        Q215*, S217V, T218A, T218L, T218Q, T218R, T218V, S221N, G222D,        E224A, E224P, S225G, N227A, N227Q, N227R, N227S, N227T, N227K,        D230*, D230N, D230R, D230T, D230W, E232D, E232V, N233D, N233E,        N233H, N233Q, N233R, N233W, W234R, G235W, G235A, G235E, G235F,        G235H, G235I, G235L, G235M, G235N, G235P, G235S, G235V, S237C,        S237G, S237M, S237N, S237W, S237Y, Y244C, Y244E, Y244M, L249H,        L249K, L249Q, L249R, L249W, L249Y, S251A, S251L, S251N, S251R,        S251W, N252P, N252C, G253D, G253W, F254I, F254L, F254M, F254Y,        Q256D, Q256E, Q256R, N260*, N260A, N260C, N260E, N260I, N260K,        N260L, N260M, N260Q, N260R, N260T, N260V, N260W, N260Y, E261*,        E261A, E261D, E261R, E261W, Q262*, Q262F, Q262H, Q262W, Q262Y,        M263K, M263L, M263Q, D264*, D264C, D264E, D264N, Y265F, N267S,        N267T, W268C, W268E, W268M, W268R, Y270F, A271D, A271G, H272D,        H272I, H272M, H272P, H272V, H272W, N273W, K274R, K274A, K274H,        F276A, F276C, F276K, F276N, F276G, F276L, F276M, F276P, F276S,        F276V, F276W, I278A, I278K, I278N, I278Q, I278V, S279C, S279D,        S279E, S279G, S279N, D280C, D280E, Y281*, Y281A, Y281C, Y281H,        Y281K, Y281N, Y281P, Y281R, Y282E, Y282N, H283I, A284I, A284L,        A284N, A284P, A284T, A284V, T287N, S288P, S288D, S288K, S288N,        V290I, K291L, K291R, K291V, T296C, E300A, E300D, H301C, H301N,        H301R, T303A, T303C, T303G, T303K, T303Q, T303R, T303W, D304C,        D304M, L305M, L305N, S306C, K308A, K308D, K308E, K308G, K308I,        K308L, K308Q, K308S, K308T, K308V, K308Y, K309A, K309C, K309D,        K309E, K309G, K309H, K309L, K309M, K309N, K309Q, K309S, K309T,        K309I, K312A, K312E, K312L, K312M, K312N, K312Q, K312S, K312W,        E314I, E314L, E314V, L315I, L315V, R319A, Y321F and N323R; and    -   b) recovering the variant, wherein the variant has        hexosaminidase, preferably beta-1,6 N-acetylglucosaminidase        activity.

One aspect of the invention relates to method for obtaining a dispersinvariant, comprising;

-   -   a) introducing into a parent dispersin one or more substitutions        selected from the group consisting of: D2V, Q3F, Q3I, S12A,        H15Y, T17W, T17E, V18L, K22M, S23I, S23C, S23A, S23T, S23L,        S23V, S23E, L24I, V25R, Y30A, Y30L, A49W, A49Y, S56T, N59D,        N59C, N59E, N59R, N59F, N59W, N59V, N62C, N62D, T63C, N68Q,        S72D, S72E, I74L, S77A, I82V, L90F, E99Q, L100S, T114S, T114C,        Y123W, Y124I, Y124M, Y124H, Y124R, Y124V, Y124Q, Y124T, Y124K,        D125R, D125C, D125G, D125H, D125K, D125Q, Q135M, D138R, D138K,        D138N, D138Q, V140I, S163P, L170D, L170K, L170S, L170H, D171E,        D171K, D171Y, D171M, D171Q, D171L, I173C, D174W, D174H, D174M,        D174N, F175Y, Q178K, I179T, S181T, S181F, S181Q, S181G, S181N,        S181C, S181K, E185A, E185R, E185M, E185V, S186K, S186M, S186R,        S186H, K187G, Y188P, E189V, S199C, S199L, A203G, A203E, A203V,        N204L, N204Y, N204V, L205I, D207N, D207S, D207C, D207G, S210T,        Q215K, Q215R, T218Q, S221N, S225G, N227T, N227K, E232D, G235W,        S237W, Y244M, Y244C, N252P, N252C, Q256E, Q256D, N260Q, Q262H,        M263Q, D264E, Y265F, N267T, N267S, Y270F, H272M, H272P, H272I,        H272V, N273I, N273W, K274H, F276A, F276N, F276K, F276C, I278V,        S279N, S279D, S279G, D280E, D280C, Y281P, Y282F, Y282N, H283I,        A284T, A284L, A284I, S288P, S288K, V290I, K291R, T296C, T303Q,        D304M, D304C, L305M, L305N, S306C, K308D, K308E, K308A, K308V,        K308Q, K308S, K308Y, K308G, K308L, K308T, K308I, K309E, K309G,        K309C, K309L, K309D, K309Q, K309N, K309T, K309A, K309S, K309M,        K309H, K312E, K312A, K312Q, K312S, K312W, K312L, K312N, E314L,        E314V, E314I, L315I, R319A, Y321F, and N323R; and    -   b) recovering the variant, wherein the variant has        hexosaminidase, preferably beta-1,6 N-acetylglucosaminidase        activity.

The method preferably comprises introduction of 2-20, e.g. 2-10 and 2-5,such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19or 20 alterations compared to the polypeptide shown in SEQ ID NO: 1.

Some aspects of the invention relate to a method for obtaining adispersin variant, comprising introducing into a parent dispersin analteration at one or more positions, wherein the dispersin parentbelongs to the Terribacillus clade and wherein the parent dispersincomprises one or more of the motifs: GXDE (SEQ ID NO:8),[EQ][NRSHA][YVFL][AGSTC][IVLF][EAQYN][SN] (SEQ ID NO:9),[VIM][LIV]G[GAV]DE[VI][PSA] (SEQ ID NO: 10), WND[SQR][IVL][TLVM] (SEQ IDNO: 11), QSTL (SEQ ID NO: 12), NKFFY (SEQ ID NO: 13) or NLD[DR]S (SEQ IDNO: 14).

Some aspects of the invention relate to a method for obtaining adispersin variant, comprising introducing into a parent dispersin analteration at one or more positions, wherein the parent dispersin isselected from the group of polypeptides:

-   -   a) a polypeptide having at least 60%, at least 65%, at least        70%, at least 75%, at least 80%, at least 85%, at least 90%, at        least 91%, at least 92%, at least 93%, at least 94%, at least        95%, at least 96%, at least 97%, at least 98%, at least 99% or        100% sequence identity to the polypeptide shown in SEQ ID NO: 1,    -   b) a polypeptide having at least 60%, at least 65%, at least        70%, at least 75%, at least 80%, at least 85%, at least 90%, at        least 91%, at least 92%, at least 93%, at least 94%, at least        95%, at least 96%, at least 97%, at least 98%, at least 99% or        100% sequence identity to the polypeptide shown in SEQ ID NO: 2,    -   c) a polypeptide having at least 60%, at least 65%, at least        70%, at least 75%, at least 80%, at least 85%, at least 90%, at        least 91%, at least 92%, at least 93%, at least 94%, at least        95%, at least 96%, at least 97%, at least 98%, at least 99% or        100% sequence identity to the polypeptide shown in SEQ ID NO: 3,    -   d) a polypeptide having at least 60%, at least 65%, at least        70%, at least 75%, at least 80%, at least 85%, at least 90%, at        least 91%, at least 92%, at least 93%, at least 94%, at least        95%, at least 96%, at least 97%, at least 98%, at least 99% or        100% sequence identity to the polypeptide shown in SEQ ID NO: 4,    -   e) a polypeptide having at least 60%, at least 65%, at least        70%, at least 75%, at least 80%, at least 85%, at least 90%, at        least 91%, at least 92%, at least 93%, at least 94%, at least        95%, at least 96%, at least 97%, at least 98%, at least 99% or        100% sequence identity to the polypeptide shown in SEQ ID NO: 5,

Some aspect relates to a method of obtaining a dispersin variant whereinthe parent dispersin is obtained from Terribacillus genus.

Some aspect relates to a method of obtaining a dispersin variant whereinthe dispersin parent has at least 60%, at least 65%, at least 70%, atleast 75%, at least 80%, at least 85%, at least 90%, at least 91%, atleast 92%, at least 93%, at least 94%, at least 95%, at least 96%, atleast 97%, at least 98%, at least 99% or 100% sequence identity to thepolypeptide shown in SEQ ID NO: 1.

The variants of the invention may be prepared by procedures such asthose mentioned below.

Site-directed mutagenesis is a technique in which one or more (e.g.,several) mutations are introduced at one or more defined sites in apolynucleotide encoding the parent.

Site-directed mutagenesis can be accomplished in vitro by PCR involvingthe use of oligonucleotide primers containing the desired mutation.Site-directed mutagenesis can also be performed in vitro by cassettemutagenesis involving the cleavage by a restriction enzyme at a site inthe plasmid comprising a polynucleotide encoding the parent andsubsequent ligation of an oligonucleotide containing the mutation in thepolynucleotide. Usually the restriction enzyme that digests the plasmidand the oligonucleotide is the same, permitting sticky ends of theplasmid and the insert to ligate to one another. See, e.g., Scherer andDavis, 1979, Proc. Natl. Acad. Sci. USA 76: 4949-4955; and Barton etal., 1990, Nucleic Acids Res. 18: 7349-4966. Site-directed mutagenesiscan also be accomplished in vivo by methods known in the art.

See, e.g., U.S. Patent Application Publication No. 2004/0171154; Storiciet al., 2001, Nature Biotechnol. 19: 773-776; Kren et al., 1998, Nat.Med. 4: 285-290; and Calissano and Macino, 1996, Fungal Genet. Newslett.43: 15-16.

Synthetic gene construction entails in vitro synthesis of a designedpolynucleotide molecule to encode a polypeptide of interest. Genesynthesis can be performed utilizing a number of techniques, such as themultiplex microchip-based technology described by Tian et al. (2004,Nature 432: 1050-1054) and similar technologies wherein oligonucleotidesare synthesized and assembled upon photo-programmable microfluidicchips.

Single or multiple amino acid substitutions, deletions, and/orinsertions can be made and tested using known methods of mutagenesis,recombination, and/or shuffling, followed by a relevant screeningprocedure, such as those disclosed by Reidhaar-Olson and Sauer, 1988,Science 241: 53-57; Bowie and Sauer, 1989, Proc. Natl. Acad. Sci. USA86: 2152-2156; WO 95/17413; or WO 95/22625. Other methods that can beused include error-prone PCR, phage display (e.g., Lowman et al., 1991,Biochemistry 30: 10832-10837; U.S. Pat. No. 5,223,409; WO 92/06204) andregion-directed mutagenesis (Derbyshire et al., 1986, Gene 46: 145; Neret al., 1988, DNA 7: 127).

Mutagenesis/shuffling methods can be combined with high-throughput,automated screening methods to detect activity of cloned, mutagenizedpolypeptides expressed by host cells (Ness et al., 1999, NatureBiotechnology 17: 893-896). Mutagenized DNA molecules that encode activepolypeptides can be recovered from the host cells and rapidly sequencedusing standard methods in the art. These methods allow the rapiddetermination of the importance of individual amino acid residues in apolypeptide.

Semi-synthetic gene construction is accomplished by combining aspects ofsynthetic gene construction, and/or site-directed mutagenesis, and/orrandom mutagenesis, and/or shuffling. Semi-synthetic construction istypified by a process utilizing polynucleotide fragments that aresynthesized, in combination with PCR techniques. Defined regions ofgenes may thus be synthesized de novo, while other regions may beamplified using site-specific mutagenic primers, while yet other regionsmay be subjected to error-prone PCR or non-error prone PCRamplification. Polynucleotide subsequences may then be shuffled.

The variants of the applications may be prepared and identified by sitesaturated library methods, such described in: “Methods and applicationsin protein engineering”, Rodrigo M. P. Siloto, Randall J. Weselake.Biocatalysis and Agricultural Biotechnology 1 (2012) 181-189.

Stabilization of an enzyme may entail running a site saturation library(SSL) screening campaign. A SSL library allows all 20 amino acids at thegiven amino acid position and a library is normally constructed for eachamino acid position in the protein. The number of SSL libraries are thustypically counted in the hundreds for a typical enzyme molecule. Theentire campaign tends to be very time consuming and resource demanding.The process will typically require construction of libraries, plating,colony picking, sequencing, re-gridding and high-throughput screening(HTS).

The screening itself is based on the principle that an exponential decayis observed for degradation of a protein when exposed to stressconditions, e.g. elevated temperature, proteases, surfactant. Astability improved variant can be found in a screening situationregardless of its initial concentration, as the stable variant aftersufficient stress will always surpass the concentration of a referencesample by comparison, given a sufficiently sensitive detection method isavailable. The concentration may be reflected in e.g. an activitymeasurement.

Surprisingly, the principle of screening described above cansuccessfully be extended to screening multiple variants for increasedhalf-life in one go, i.e. screened as a single library pool or a mixtureof variants. The first round of screening will not identify thestabilizing substitution itself, but it will enable a prioritization ofthe many libraries according to likelihood of containing variant(s) ofinterest based on selected parameters (e.g. thermostability, stabilityin presence of surfactant).

The result of screening could for example be the level of stabilitydescribed by the ratio in activity between a stressed and an unstressedlibrary. The actual measurement of activity in a stressed pool (and thebackbone reference), can be conducted in several ways, spanning from asingle end-of-stress activity measurement to multiple measurementscharacterizing the degradation as a function of time or increasingstress. The presence of improved variant(s) in a library will becomeincreasing profound as the pool (and backbone reference) becomesincreasingly degraded. It is therefore advantageous to measure the levelof degradation late in the stress process.

Libraries with the highest level of stability, and preferably above thatdetermined for the reference, are selected to be most interesting. Theresult obtained for a library is the combined effect of all variants itcontains and the likelihood of finding improved variants in a specificlibrary therefore increases with the level of its overall stability.Several factors can contribute to a positive screening result for alibrary, e.g. small sized library, variants having a high half-life,high concentration of stable variants, sensitive detection method toallow high stress conditions. The variants contained in the libraries ofthe selected amino acid positions are then tested for improvedproperties as isolated clones. The number of positions selected may inprinciple range from 1 to the total number of amino acid positions, butit will in practice always be significantly lower than the total numberof positions.

In summary, the method of screening mixtures of variants is particularlyuseful in the context of screening small libraries targeting a singleposition in the context of protein engineering, e.g. libraries generatedusing NNS or other degenerate nucleotides at a given codon. Thescreening principle described can be carried out for multiple differentsmall libraries, each representing e.g. full or partial site saturationof a single position, thus investigating multiple positions in parallel.This enables a quick assessment of the potential for hits in multiplepositions in a single step. Promising positions can be prioritized andfurther investigated, leading to faster identification of hits(stabilizing substitutions). In summary, the method saves time, labourand consumables by significantly reducing the number of clones requiredto be screened for identification of stabilizing substitutions.

The method of the invention comprises; preparing a library ofpolypeptide variants (V_(L)), for each position (P) in the polypeptide.Up to all the 20 possible amino acids are introduced by substitutionresulting in a specific number of different variants for each position.This can be done by well-known methods of generating diversity in thepositions of the polypeptide. Some examples include but are not limitedto NNS doping, where the codons comprise all four possible nucleobases A(adenine), C (cytosine), G (guanine), T (thymine) at position 1 and 2,whereas the third position comprises C (cytosine), G (guanine) only. Inthat way codons encoding all 20 natural amino acids are introducedrandomly. The method diverges from traditional way of screeningindividual clones from the library: The inventors have found that thelibrary V_(L) could be subjected to measurement directly and from thismeasurement, libraries including improved variants could be selecteddirectly. This enables selecting a subset library (V_(LSub)) containingimproved variants. This subset V_(LSub) is subjected to steps 1 to 4:

-   -   1. plate clones on agar plates,    -   2. pick each colony grown on the plates,    -   3. grow the picked individual colonies in microtiter plates, and    -   4. screen and sequence grown colonies        Unlike the normal procedure these four steps only need to be        performed for selected subsets and not for each position e.g.        324 times for a polypeptide having 324 positions. The steps 1-4        are very time consuming. The method of the current invention        therefore significantly reduces the amount of time for making        improved variants by limiting the number of libraries processed        in steps 1 to 4.

One embodiment of the invention relates to a method for finding improvedpolypeptide variants, comprising the steps of:

-   -   a) generate diversity in selected, preferably all, positions of        a polypeptide, thereby preparing libraries of polypeptide        variants (V_(L)), and    -   b) select subset library (V_(LSub)) with improved variants.        The subset library (V_(LSub)) may be selected based on different        criteria such as improved stability, activity etc. as already        described above. Thus, in one embodiment the libraries of        polypeptide variants V_(L) are subjected to a stress condition,        e.g. elevated temperature, and a subset library (V_(LSub)) is        selected from V_(L).

One embodiment of the invention relates to a method for finding improvedpolypeptide variants, comprising the steps of:

-   -   a) generate diversity in selected, preferably all, positions of        a polypeptide, thereby preparing a library of polypeptide        variants (V_(L)),    -   b) expose part of the library of polypeptide variants (V_(L)) to        one or more stress conditions suitable for measuring a property        of interest, thereby generating a stressed V_(LS+n) and an        unstressed V_(LUS) library;    -   c) compare the stressed V_(LS) and the unstressed V_(LUS)        library; and    -   d) select subset library (V_(LSub)) with improved variants.        Where V_(LS+n) reflects that any number of library stressed        subset could be made i.e. n≥1. Preferably, the subset library is        subjected to the steps 1 to 4 as described above. Thus, one        embodiment of the invention relates to a method for finding        improved polypeptide variants, comprising the steps of:

a) generate diversity in selected, preferably all, positions of apolypeptide, thereby preparing a library of polypeptide variants(V_(L)), and

-   -   b) select subset library (V_(LSub)) with improved variants,    -   c) plate selected subset library (V_(LSub)) clones on agar        plates,    -   d) pick each colony grown on the plates,    -   e) grow the picked individual colonies e.g. in microtiter plates    -   f) select individual improved variants by screening, and    -   g) sequence selected variants to identify specific mutations.        One embodiment of the invention relates to a method for finding        improved polypeptide variants, comprising the steps of:    -   a) generate diversity in selected, preferably all, positions of        a polypeptide, thereby preparing a library of polypeptide        variants (V_(L)),    -   b) grow the individual libraries e.g. in microtiter plates    -   c) expose part of the library of polypeptide variants (V_(L)) to        one or more stress conditions suitable for measuring a property        of interest, thereby generating a stressed V_(LS+n) and an        unstressed V_(LUS) library;    -   d) compare the stressed V_(LS) and the unstressed V_(LUS)        library; and    -   e) select subset library (V_(LSub)) with improved variants    -   f) plate selected subset library (V_(LSub)) clones on agar        plates,    -   g) pick each colony grown on the plates,    -   h) grow the picked individual colonies e.g. in microtiter plates    -   i) select individual improved variants by screening, and    -   j) sequence selected variants to identify specific mutations.        In one embodiment the number of variants in the subset library        (V_(LSub)) is in the range of 2-20 polypeptide variants,        preferable in the range of 5-20 polypeptide variants, preferable        in the range of 5-10 polypeptide variants, preferable in the        range of 8-20 polypeptide variants or in the range of 8-15        polypeptide variants.

In one embodiment, the library of polypeptide variants (V_(L)) isgenerated by using doped primers e.g. using NNS doping as describedabove. In one embodiment, the library of polypeptide variants (V_(L)) isgenerated by using doped primers e.g. using NMS, NKS, NRS, NYS, NWS,NSS, MNS, KNS, RNS, YNS, WNS, SNS, NAS, NCS, NGS, NTS, ANS, CNS, GNS,TNS, NNK, RNS, YHS, AYS, WRS (IUPAC nucleotide codes) doping asdescribed above

In one embodiment, the library (V_(L)) is sequenced to confirmdiversity. The distribution of codon usage in the library may bedetermined by comparison of sequence reads covering the targetedposition using any of the methods known in the art e.g. such as nextgeneration sequencing (NGS) as described in Ravi R. K., Walton K.,Khosroheidari M. (2018) MiSeq: A Next Generation Sequencing Platform forGenomic Analysis. In: DiStefano J. (eds) Disease Gene Identification.Methods in Molecular Biology, vol 1706. Humana Press, New York, N.Y.

The polypeptide is preferably an enzyme but may be applied to anyprotein. The enzyme may be selected from any suitable enzymes such ashydrolase, isomerase, ligase, lyase, oxidoreductase, or transferase,e.g., an alpha-galactosidase, alpha-glucosidase, aminopeptidase,amylase, beta-galactosidase, beta-glucosidase, beta-xylosidase,carbohydrase, carboxypeptidase, catalase, cellobiohydrolase, cellulase,chitinase, cutinase, cyclodextrin glycosyltransferase,deoxyribonuclease, endoglucanase, esterase, glucoamylase,hexosaminidases e.g. dispersins, invertase, laccase, esterases, lipase,mannosidase, mannanases, mutanase, nucleases e.g. DNases or RNases,oxidase, pectate lyase, pectinolytic enzyme, peroxidase, phytase,polyphenoloxidase, proteolytic enzyme proteases, transglutaminase andxylanase. The enzyme is prefereably a dispersin and preferably themixture of variants are variants of the present invention.

The property of interest may be e.g. stability, such asthermo-stability, in-wash stability, detergent stability, storagestability, secondary enzyme e.g. protease stability, sulfide-stabilityetc. all of which is not mutually exclusive. The improved property maybe performance or activity e.g. such as wash performance, increasedactivity on specific substrate. The property may also be improved oraltered substrate specificity.

One embodiment relates to a method for identifying and selectingvariants with improved stability comprising the steps of:

-   -   a) generate diversity in selected, preferably all, positions of        a polypeptide, thereby preparing a library of polypeptide        variants (V_(L)),    -   b) divide the library of polypeptide variants (V_(L)) in two,        expose one half of the library of polypeptide variants (V_(L))        to one or more stress conditions suitable for measuring        stability, thereby generating a stressed V_(LS) and an        unstressed V_(LUS) library;    -   c) compare the stressed V_(LS) and the unstressed V_(LUS)        library; and    -   d) select subset library (V_(LSub))with stability improved        variants.        The stability is preferably measured as Half-life improvement        factor (HIF) or residual activity ratio (RAR) as described in        the “dispersin variants” section above. The stress condition is        preferably elevated temperature such as above 37° C., above 41°        C., above 46° C. or above 50° C.        The property of interest may also be activity defined as        improved substrate turn over. In one embodiment the property of        interest is improved dispersin activity, such as increased        catalysis of β-1,6-glycosidic linkages of N-acetyl-glucosamine        polymers (poly-N-acetylglucosamine, PNAG) i.e. the dispersin        variant display improved activity relative (or compared) to the        activity of the parent dispersin, such as compared to a        dispersin with SEQ ID NO: 1 or compared to a dispersin having        the identical amino acid sequence of said variant but not having        the alterations at one or more of said specified positions.        One embodiment relates to a method for identifying and selecting        variants with improved beta-1,6 N-acetylglucosaminidase activity        comprising the steps of    -   a) generate diversity in selected, preferably all, positions of        a polypeptide, thereby preparing a library of polypeptide        variants (V_(L)),    -   b) divide the library of polypeptide variants (V_(L)) in two,        expose one half of the library of polypeptide variants (V_(L))        to one or more stress conditions suitable for measuring beta-1,6        N-acetylglucosaminidase activity, thereby generating a stressed        V_(LS) and an unstressed V_(LUS) library;    -   c) compare the stressed V_(LS) and the unstressed V_(LUS)        library; and    -   d) select subset library (V_(LSub)) with beta-1,6        N-acetylglucosaminidase activity improved variants. It is clear        to the skilled artisan that the term improved refers to the        polypeptide variants being improved compared to a reference        polypeptide, which may be the parent polypeptide or a        polypeptide with SEQ ID NO: 1.

Nucleic Acid Constructs

The present invention also relates to nucleic acid constructs comprisinga polynucleotide encoding the dispersin variants of the presentinvention operably linked to one or more control sequences that directthe expression of the coding sequence in a suitable host cell underconditions compatible with the control sequences.

The polynucleotide may be manipulated in a variety of ways to providefor expression of the polypeptide. Manipulation of the polynucleotideprior to its insertion into a vector may be desirable or necessarydepending on the expression vector. The techniques for modifyingpolynucleotides utilizing recombinant DNA methods are well known in theart.

The control sequence may be a promoter, a polynucleotide that isrecognized by a host cell for expression of a polynucleotide encoding apolypeptide of the present invention. The promoter containstranscriptional control sequences that mediate the expression of thepolypeptide. The promoter may be any polynucleotide that showstranscriptional activity in the host cell including mutant, truncated,and hybrid promoters, and may be obtained from genes encodingextracellular or intracellular polypeptides either homologous orheterologous to the host cell.

Examples of suitable promoters for directing transcription of thenucleic acid constructs of the present invention in a bacterial hostcell are the promoters obtained from the Bacillus amyloliquefaciensalpha-amylase gene (amyQ), Bacillus licheniformis alpha-amylase gene(amyL), Bacillus licheniformis penicillinase gene (penP), Bacillusstearothermophilus maltogenic amylase gene (amyM), Bacillus subtilislevansucrase gene (sacB), Bacillus subtilis xylA and xylB genes,Bacillus thuringiensis cryllIA gene (Agaisse and Lereclus, 1994,Molecular Microbiology 13: 97-107), E. coli lac operon, E. coli trcpromoter (Egon et al., 1988, Gene 69: 301-315), Streptomyces coelicoloragarase gene (dagA), and prokaryotic beta-lactamase gene (Villa-Kamaroffet al., 1978, Proc. Natl. Acad. Sci. USA 75: 3727-3731), as well as thetac promoter (DeBoer et al., 1983, Proc. Natl. Acad. Sci. USA 80:21-25). Further promoters are described in “Useful proteins fromrecombinant bacteria” in Gilbert et al., 1980, Scientific American 242:74-94; and in Sambrook et al., 1989, supra. Examples of tandem promotersare disclosed in WO 99/43835.

Examples of suitable promoters for directing transcription of thepolynucleotide of the present invention in a filamentous fungal hostcell are promoters obtained from the genes for Aspergillus nidulansacetamidase, Aspergillus niger neutral alpha-amylase, Aspergillus nigeracid stable alpha-amylase, Aspergillus niger or Aspergillus awamoriglucoamylase (glaA), Aspergillus oryzae TAKA amylase, Aspergillus oryzaealkaline protease, Aspergillus oryzae triose phosphate isomerase,Fusarium oxysporum trypsin-like protease (WO 96/00787), Fusariumvenenatum amyloglucosidase (WO 00/56900), Fusarium venenatum Daria (WO00/56900), Fusarium venenatum Quinn (WO 00/56900), Rhizomucor mieheilipase, Rhizomucor miehei aspartic proteinase, Trichoderma reeseibeta-glucosidase, Trichoderma reesei cellobiohydrolase I, Trichodermareesei cellobiohydrolase II, Trichoderma reesei endoglucanase I,Trichoderma reesei endoglucanase II, Trichoderma reesei endoglucanaseIII, Trichoderma reesei endoglucanase V, Trichoderma reesei xylanase I,Trichoderma reesei xylanase II, Trichoderma reesei xylanase III,Trichoderma reesei beta-xylosidase, and Trichoderma reesei translationelongation factor, as well as the NA2-tpi promoter (a modified promoterfrom an Aspergillus neutral alpha-amylase gene in which the untranslatedleader has been replaced by an untranslated leader from an Aspergillustriose phosphate isomerase gene; non-limiting examples include modifiedpromoters from an Aspergillus niger neutral alpha-amylase gene in whichthe untranslated leader has been replaced by an untranslated leader froman Aspergillus nidulans or Aspergillus oryzae those phosphate isomerasegene); and mutant, truncated, and hybrid promoters thereof. Otherpromoters are described in U.S. Pat. No. 6,011,147.

In a yeast host, useful promoters are obtained from the genes forSaccharomyces cerevisiae enolase (ENO: 1), Saccharomyces cerevisiaegalactokinase (GAL1), Saccharomyces cerevisiae alcoholdehydrogenase/glyceraldehyde-3 phosphate dehydrogenase (ADH1, ADH2/GAP),Saccharomyces cerevisiae triose phosphate isomerase (TPI), Saccharomycescerevisiae metallothionein (CUP1), and Saccharomyces cerevisiae 3phosphoglycerate kinase. Other useful promoters for yeast host cells aredescribed by Romanos et al., 1992, Yeast 8: 423-488.

The control sequence may also be a transcription terminator, which isrecognized by a host cell to terminate transcription. The terminator isoperably linked to the 3′ terminus of the polynucleotide encoding thepolypeptide. Any terminator that is functional in the host cell may beused in the present invention.

Preferred terminators for bacterial host cells are obtained from thegenes for Bacillus clausii alkaline protease (aprH), Bacilluslicheniformis alpha-amylase (amyL), and Escherichia coli ribosomal RNA(rrnB).

Preferred terminators for filamentous fungal host cells are obtainedfrom the genes for Aspergillus nidulans acetamidase, Aspergillusnidulans anthranilate synthase, Aspergillus niger glucoamylase,Aspergillus niger alpha-glucosidase, Aspergillus oryzae TAKA amylase,Fusarium oxysporum trypsin-like protease, Trichoderma reeseibeta-glucosidase, Trichoderma reesei cellobiohydrolase I, Trichodermareesei cellobiohydrolase II, Trichoderma reesei endoglucanase I,Trichoderma reesei endoglucanase II, Trichoderma reesei endoglucanaseIII, Trichoderma reesei endoglucanase V, Trichoderma reesei xylanase I,Trichoderma reesei xylanase II, Trichoderma reesei xylanase Ill,Trichoderma reesei beta-xylosidase, and Trichoderma reesei translationelongation factor.

Preferred terminators for yeast host cells are obtained from the genesfor Saccharomyces cerevisiae enolase, Saccharomyces cerevisiaecytochrome C (CYC1), and Saccharomyces cerevisiae glyceraldehyde-3phosphate dehydrogenase. Other useful terminators for yeast host cellsare described by Romanos et al., 1992, supra.

The control sequence may also be an mRNA stabilizer region downstream ofa promoter and upstream of the coding sequence of a gene which increasesexpression of the gene.

Examples of suitable mRNA stabilizer regions are obtained from aBacillus thuringiensis ctylllA gene (WO 94/25612) and a Bacillussubtilis SP82 gene (Hue et al., 1995, J. Bacteriol. 177: 3465-3471).

The control sequence may also be a leader, a nontranslated region of anmRNA that is important for translation by the host cell. The leader isoperably linked to the 5′ terminus of the polynucleotide encoding thepolypeptide. Any leader that is functional in the host cell may be used.

Preferred leaders for filamentous fungal host cells are obtained fromthe genes for Aspergillus oryzae TAKA amylase and Aspergillus nidulanstriose phosphate isomerase.

Suitable leaders for yeast host cells are obtained from the genes forSaccharomyces cerevisiae enolase (ENO-1), Saccharomyces cerevisiae3-phosphoglycerate kinase, Saccharomyces cerevisiae alpha-factor, andSaccharomyces cerevisiae alcoholdehydrogenase/glyceraldehyde-3-phosphate dehydrogenase (ADH2/GAP).

The control sequence may also be a polyadenylation sequence, a sequenceoperably linked to the 3′ terminus of the polynucleotide and, whentranscribed, is recognized by the host cell as a signal to addpolyadenosine residues to transcribed mRNA. Any polyadenylation sequencethat is functional in the host cell may be used.

Preferred polyadenylation sequences for filamentous fungal host cellsare obtained from the genes for Aspergillus nidulans anthranilatesynthase, Aspergillus niger glucoamylase, Aspergillus nigeralpha-glucosidase, Aspergillus oryzae TAKA amylase, and Fusariumoxysporum trypsin-like protease.

Useful polyadenylation sequences for yeast host cells are described byGuo and Sherman, 1995, Mol. Cellular Biol. 15: 5983-5990.

The control sequence may also be a signal peptide coding region thatencodes a signal peptide linked to the N terminus of a polypeptide anddirects the polypeptide into the cell's secretory pathway. The 5′ end ofthe coding sequence of the polynucleotide may inherently contain asignal peptide coding sequence naturally linked in translation readingframe with the segment of the coding sequence that encodes thepolypeptide. Alternatively, the 5′ end of the coding sequence maycontain a signal peptide coding sequence that is foreign to the codingsequence. A foreign signal peptide coding sequence may be required wherethe coding sequence does not naturally contain a signal peptide codingsequence. Alternatively, a foreign signal peptide coding sequence maysimply replace the natural signal peptide coding sequence in order toenhance secretion of the polypeptide. However, any signal peptide codingsequence that directs the expressed polypeptide into the secretorypathway of a host cell may be used.

Effective signal peptide coding sequences for bacterial host cells arethe signal peptide coding sequences obtained from the genes for BacillusNCIB 11837 maltogenic amylase, Bacillus licheniformis subtilisin,Bacillus licheniformis beta-lactamase, Bacillus stearothermophilusalpha-amylase, Bacillus stearothermophilus neutral proteases (nprT,nprS, nprM), and Bacillus subtilis prsA. Further signal peptides aredescribed by Simonen and Palva, 1993, Microbiological Reviews 57:109-137.

Effective signal peptide coding sequences for filamentous fungal hostcells are the signal peptide coding sequences obtained from the genesfor Aspergillus niger neutral amylase, Aspergillus niger glucoamylase,Aspergillus oryzae TAKA amylase, Humicola insolens cellulase, Humicolainsolens endoglucanase V, Humicola lanuginosa lipase, and Rhizomucormiehei aspartic proteinase.

Useful signal peptides for yeast host cells are obtained from the genesfor Saccharomyces cerevisiae alpha-factor and Saccharomyces cerevisiaeinvertase. Other useful signal peptide coding sequences are described byRomanos et al., 1992, supra.

The control sequence may also be a propeptide coding sequence thatencodes a propeptide positioned at the N terminus of a polypeptide. Theresultant polypeptide is known as a proenzyme or propolypeptide (or azymogen in some cases). A propolypeptide is generally inactive and canbe converted to an active polypeptide by catalytic or autocatalyticcleavage of the propeptide from the propolypeptide. The propeptidecoding sequence may be obtained from the genes for Bacillus subtilisalkaline protease (aprE), Bacillus subtilis neutral protease (nprT),Myceliophthora thermophila laccase (WO 95/33836), Rhizomucor mieheiaspartic proteinase, and Saccharomyces cerevisiae alpha-factor.

Where both signal peptide and propeptide sequences are present, thepropeptide sequence is positioned next to the N terminus of apolypeptide and the signal peptide sequence is positioned next to the Nterminus of the propeptide sequence.

It may also be desirable to add regulatory sequences that regulateexpression of the polypeptide relative to the growth of the host cell.Examples of regulatory sequences are those that cause expression of thegene to be turned on or off in response to a chemical or physicalstimulus, including the presence of a regulatory compound. Regulatorysequences in prokaryotic systems include the lac, tac, and trp operatorsystems. In yeast, the ADH2 system or GAL1 system may be used. Infilamentous fungi, the Aspergillus niger glucoamylase promoter,Aspergillus oryzae TAKA alpha-amylase promoter, and Aspergillus oryzaeglucoamylase promoter, Trichoderma reesei cellobiohydrolase I promoter,and Trichoderma reesei cellobiohydrolase II promoter may be used. Otherexamples of regulatory sequences are those that allow for geneamplification. In eukaryotic systems, these regulatory sequences includethe dihydrofolate reductase gene that is amplified in the presence ofmethotrexate, and the metallothionein genes that are amplified withheavy metals. In these cases, the polynucleotide encoding thepolypeptide would be operably linked to the regulatory sequence.

Expression Vectors

The present invention also relates to recombinant expression vectorscomprising a polynucleotide encoding the dispersin variants of thepresent invention, a promoter, and transcriptional and translationalstop signals. The various nucleotide and control sequences may be joinedtogether to produce a recombinant expression vector that may include oneor more convenient restriction sites to allow for insertion orsubstitution of the polynucleotide encoding the polypeptide at suchsites. Alternatively, the polynucleotide may be expressed by insertingthe polynucleotide or a nucleic acid construct comprising thepolynucleotide into an appropriate vector for expression. In creatingthe expression vector, the coding sequence is located in the vector sothat the coding sequence is operably linked with the appropriate controlsequences for expression. In a further aspect, polynucleotide sequencecodons have been modified by nucleotide substitutions to correspond tothe codon usage of the host organism intended for production of thepolypeptide of the present invention. The recombinant expression vectormay be any vector (e.g., a plasmid or virus) that can be convenientlysubjected to recombinant DNA procedures and can bring about expressionof the polynucleotide. The choice of the vector will typically depend onthe compatibility of the vector with the host cell into which the vectoris to be introduced. The vector may be a linear or closed circularplasmid.

The vector may be an autonomously replicating vector, i.e., a vectorthat exists as an extrachromosomal entity, the replication of which isindependent of chromosomal replication, e.g., a plasmid, anextrachromosomal element, a minichromosome, or an artificial chromosome.The vector may contain any means for assuring self-replication.Alternatively, the vector may be one that, when introduced into the hostcell, is integrated into the genome and replicated together with thechromosome(s) into which it has been integrated. Furthermore, a singlevector or plasmid or two or more vectors or plasmids that togethercontain the total DNA to be introduced into the genome of the host cell,or a transposon, may be used.

The vector preferably contains one or more selectable markers thatpermit easy selection of transformed, transfected, transduced, or thelike cells. A selectable marker is a gene the product of which providesfor biocide or viral resistance, resistance to heavy metals, prototrophyto auxotrophs, and the like.

Examples of bacterial selectable markers are Bacillus licheniformis orBacillus subtilis dal genes, or markers that confer antibioticresistance such as ampicillin, chloramphenicol, kanamycin, neomycin,spectinomycin, or tetracycline resistance. Suitable markers for yeasthost cells include, but are not limited to, ADE2, HIS3, LEU2, LYS2,MET3, TRP1, and URA3. Selectable markers for use in a filamentous fungalhost cell include, but are not limited to, adeA(phosphoribosylaminoimidazole-succinocarboxamide synthase), adeB(phosphoribosyl-aminoimidazole synthase), amdS (acetamidase), argB(ornithine carbamoyltransferase), bar (phosphinothricinacetyltransferase), hph (hygromycin phosphotransferase), niaD (nitratereductase), pyrG (orotidine-5′-phosphate decarboxylase), sC (sulfateadenyltransferase), and trpC (anthranilate synthase), as well asequivalents thereof. Preferred for use in an Aspergillus cell areAspergillus nidulans or Aspergillus oryzae amdS and pyrG genes and aStreptomyces hygroscopicus bar gene. Preferred for use in a Trichodermacell are adeA, adeB, amdS, hph, and pyrG genes.

The selectable marker may be a dual selectable marker system asdescribed in WO 2010/039889. In some aspects, the dual selectable markeris an hph-tk dual selectable marker system.

The vector preferably contains an element(s) that permits integration ofthe vector into the host cell's genome or autonomous replication of thevector in the cell independent of the genome. For integration into thehost cell genome, the vector may rely on the polynucleotide's sequenceencoding the polypeptide or any other element of the vector forintegration into the genome by homologous or non-homologousrecombination. Alternatively, the vector may contain additionalpolynucleotides for directing integration by homologous recombinationinto the genome of the host cell at a precise location(s) in thechromosome(s). To increase the likelihood of integration at a preciselocation, the integrational elements should contain a sufficient numberof nucleic acids, such as 100 to 10,000 base pairs, 400 to 10,000 basepairs, and 800 to 10,000 base pairs, which have a high degree ofsequence identity to the corresponding target sequence to enhance theprobability of homologous recombination. The integrational elements maybe any sequence that is homologous with the target sequence in thegenome of the host cell. Furthermore, the integrational elements may benon-encoding or encoding polynucleotides. On the other hand, the vectormay be integrated into the genome of the host cell by non-homologousrecombination.

For autonomous replication, the vector may further comprise an origin ofreplication enabling the vector to replicate autonomously in the hostcell in question. The origin of replication may be any plasmidreplicator mediating autonomous replication that functions in a cell.The term “origin of replication” or “plasmid replicator” means apolynucleotide that enables a plasmid or vector to replicate in vivo.

Examples of bacterial origins of replication are the origins ofreplication of plasmids pBR322, pUC19, pACYC177, and pACYC184 permittingreplication in E. coli, and pUB110, pE194, pTA1060, and pAMß1 permittingreplication in Bacillus.

Examples of origins of replication for use in a yeast host cell are the2 micron origin of replication, ARS1, ARS4, the combination of ARS1 andCEN3, and the combination of ARS4 and CEN6.

Examples of origins of replication useful in a filamentous fungal cellare AMA1 and ANS1 (Gems et al., 1991, Gene 98: 61-67; Cullen et al.,1987, Nucleic Acids Res. 15: 9163-9175; WO 00/24883). Isolation of theAMA1 gene and construction of plasmids or vectors comprising the genecan be accomplished according to the methods disclosed in WO 00/24883.

More than one copy of a polynucleotide of the present invention may beinserted into a host cell to increase production of a polypeptide. Anincrease in the copy number of the polynucleotide can be obtained byintegrating at least one additional copy of the sequence into the hostcell genome or by including an amplifiable selectable marker gene withthe polynucleotide where cells containing amplified copies of theselectable marker gene, and thereby additional copies of thepolynucleotide, can be selected for by cultivating the cells in thepresence of the appropriate selectable agent.

The procedures used to ligate the elements described above to constructthe recombinant expression vectors of the present invention are wellknown to one skilled in the art (see, e.g., Sambrook et al., 1989,supra).

Host Cells

The present invention also relates to recombinant host cells, comprisinga polynucleotide of the present invention operably linked to one or morecontrol sequences that direct the production of a polypeptide of thepresent invention. A construct or vector comprising a polynucleotide isintroduced into a host cell so that the construct or vector ismaintained as a chromosomal integrant or as a self-replicatingextra-chromosomal vector as described earlier. The term “host cell”encompasses any progeny of a parent cell that is not identical to theparent cell due to mutations that occur during replication. The choiceof a host cell will to a large extent depend upon the gene encoding thepolypeptide and its source.

The host cell may be any cell useful in the recombinant production of apolypeptide of the present invention, e.g., a prokaryote or a eukaryote.

The prokaryotic host cell may be any Gram-positive or Gram-negativebacterium. Gram-positive bacteria include, but are not limited to,Bacillus, Clostridium, Enterococcus, Geobacillus, Lactobacillus,Lactococcus, Oceanobacillus, Staphylococcus, Streptococcus, andStreptomyces. Gram-negative bacteria include, but are not limited to,Campylobacter, E. coli, Flavobacterium, Fusobacterium, Helicobacter,llyobacter, Neisseria, Pseudomonas, Salmonella, and Ureaplasma.

The bacterial host cell may be any Bacillus cell including, but notlimited to, Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillusbrevis, Bacillus circulans, Bacillus clausii, Bacillus coagulans,Bacillus firmus, Bacillus lautus, Bacillus lentus, Bacilluslicheniformis, Bacillus megaterium, Bacillus pumilus, Bacillusstearothermophilus, Bacillus subtilis, and Bacillus thuringiensis cells.

The bacterial host cell may also be any Streptococcus cell including,but not limited to, Streptococcus equisimilis, Streptococcus pyogenes,Streptococcus uberis, and Streptococcus equi subsp. Zooepidemicus cells.

The bacterial host cell may also be any Streptomyces cell including, butnot limited to, Streptomyces achromogenes, Streptomyces avermitilis,Streptomyces coelicolor, Streptomyces griseus, and Streptomyces lividanscells.

The introduction of DNA into a Bacillus cell may be effected byprotoplast transformation (see, e.g., Chang and Cohen, 1979, Mol. Gen.Genet. 168: 111-115), competent cell transformation (see, e.g., Youngand Spizizen, 1961, J. Bacteriol. 81: 823-829, or Dubnau andDavidoff-Abelson, 1971, J. Mol. Biol. 56: 209-221), electroporation(see, e.g., Shigekawa and Dower, 1988, Biotechniques 6: 742-751), orconjugation (see, e.g., Koehler and Thorne, 1987, J. Bacteriol. 169:5271-5278). The introduction of DNA into an E. coli cell may be effectedby protoplast transformation (see, e.g., Hanahan, 1983, J. Mol. Biol.166: 557-580) or electroporation (see, e.g., Dower et al., 1988, NucleicAcids Res. 16: 6127-6145). The introduction of DNA into a Streptomycescell may be effected by protoplast transformation, electroporation (see,e.g., Gong et al., 2004, Folia Microbiol. (Praha) 49: 399-405),conjugation (see, e.g., Mazodier et al., 1989, J. Bacteriol. 171:3583-3585), or transduction (see, e.g., Burke et al., 2001, Proc. Natl.Acad. Sci. USA 98: 6289-6294). The introduction of DNA into aPseudomonas cell may be effected by electroporation (see, e.g., Choi etal., 2006, J. Microbiol. Methods 64: 391-397) or conjugation (see, e.g.,Pinedo and Smets, 2005, Appl. Environ. Microbiol. 71: 51-57). Theintroduction of DNA into a Streptococcus cell may be effected by naturalcompetence (see, e.g., Perry and Kuramitsu, 1981, Infect. Immun. 32:1295-1297), protoplast transformation (see, e.g., Catt and Jollick,1991, Microbios 68: 189-207), electroporation (see, e.g., Buckley etal., 1999, Appl. Environ. Microbiol. 65: 3800-3804), or conjugation(see, e.g., Clewell, 1981, Microbiol. Rev. 45: 409-436). However, anymethod known in the art for introducing DNA into a host cell can beused.

The host cell may also be a eukaryote, such as a mammalian, insect,plant, or fungal cell.

The host cell may be a fungal cell. “Fungi” as used herein includes thephyla Ascomycota, Basidiomycota, Chytridiomycota, and Zygomycota as wellas the Oomycota and all mitosporic fungi (as defined by Hawksworth etal., In, Ainsworth and Bisby's Dictionary of The Fungi, 8th edition,1995, CAB International, University Press, Cambridge, UK).

The fungal host cell may be a yeast cell. “Yeast” as used hereinincludes ascosporogenous yeast (Endomycetales), basidiosporogenousyeast, and yeast belonging to the Fungi Imperfecti (Blastomycetes).Since the classification of yeast may change in the future, for thepurposes of this invention, yeast shall be defined as described inBiology and Activities of Yeast (Skinner, Passmore, and Davenport,editors, Soc. App. Bacteriol. Symposium Series No. 9, 1980).

The yeast host cell may be a Candida, Hansenula, Kluyveromyces, Pichia,Saccharomyces, Schizosaccharomyces, or Yarrowia cell, such as aKluyveromyces lactis, Saccharomyces carlsbergensis, Saccharomycescerevisiae, Saccharomyces diastaticus, Saccharomyces douglasii,Saccharomyces kluyveri, Saccharomyces norbensis, Saccharomycesoviformis, or Yarrowia lipolytica cell.

The fungal host cell may be a filamentous fungal cell. “Filamentousfungi” include all filamentous forms of the subdivision Eumycota andOomycota (as defined by Hawksworth et al., 1995, supra). The filamentousfungi are generally characterized by a mycelial wall composed of chitin,cellulose, glucan, chitosan, mannan, and other complex polysaccharides.Vegetative growth is by hyphal elongation and carbon catabolism isobligately aerobic. In contrast, vegetative growth by yeasts such asSaccharomyces cerevisiae is by budding of a unicellular thallus andcarbon catabolism may be fermentative.

The filamentous fungal host cell may be an Acremonium, Aspergillus,Aureobasidium, Bjerkandera, Ceriporiopsis, Chrysosporium, Coprinus,Coriolus, Cryptococcus, Filibasidium, Fusarium, Humicola, Magnaporthe,Mucor, Myceliophthora, Neocallimastix, Neurospora, Paecilomyces,Penicillium, Phanerochaete, Phlebia, Piromyces, Pleurotus,Schizophyllum, Talaromyces, Thermoascus, Thielavia, Tolypocladium,Trametes, or Trichoderma cell.

For example, the filamentous fungal host cell may be an Aspergillusawamori, Aspergillus foetidus, Aspergillus fumigatus, Aspergillusjaponicus, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae,Bjerkandera adusta, Ceriporiopsis aneirina, Ceriporiopsis caregiea,Ceriporiopsis gilvescens, Ceriporiopsis pannocinta, Ceriporiopsisrivulosa, Ceriporiopsis subrufa, Ceriporiopsis subvermispora,Chrysosporium inops, Chrysosporium keratinophilum, Chrysosporiumlucknowense, Chrysosporium merdarium, Chrysosporium pannicola,Chrysosporium queenslandicum, Chrysosporium tropicum, Chrysosporiumzonatum, Coprinus cinereus, Coriolus hirsutus, Fusarium bactridioides,Fusarium cerealis, Fusarium crookwellense, Fusarium culmorum, Fusariumgraminearum, Fusarium graminum, Fusarium heterosporum, Fusarium negundi,Fusarium oxysporum, Fusarium reticulatum, Fusarium roseum, Fusariumsambucinum, Fusarium sarcochroum, Fusarium sporotrichioides, Fusariumsulphureum, Fusarium torulosum, Fusarium trichothecioides, Fusariumvenenatum, Humicola insolens, Humicola lanuginosa, Mucor miehei,Myceliophthora thermophila, Neurospora crassa, Penicillium purpurogenum,Phanerochaete chrysosporium, Phlebia radiata, Pleurotus eryngii,Talaromyces emersonii, Thielavia terrestris, Trametes villosa, Trametesversicolor, Trichoderma harzianum, Trichoderma koningii, Trichodermalongibrachiatum, Trichoderma reesei, or Trichoderma viride cell.

Fungal cells may be transformed by a process involving protoplastformation, transformation of the protoplasts, and regeneration of thecell wall in a manner known per se. Suitable procedures fortransformation of Aspergillus and Trichoderma host cells are describedin EP 238023, Yelton et al., 1984, Proc. Natl. Acad. Sci. USA 81:1470-1474, and Christensen et al., 1988, Bio/Technology 6: 1419-1422.Suitable methods for transforming Fusarium species are described byMalardier et al., 1989, Gene 78: 147-156, and WO 96/00787. Yeast may betransformed using the procedures described by Becker and Guarente, InAbelson, J.N. and Simon, M.I., editors, Guide to Yeast Genetics andMolecular Biology, Methods in Enzymology, Volume 194, pp 182-187,Academic Press, Inc., New York; Ito et al., 1983, J. Bacteriol. 153:163; and Hinnen et al., 1978, Proc. Natl. Acad. Sci. USA 75: 1920.

Methods of Production

The present invention also relates to methods of producing a dispersinvariant of the present invention, comprising (a) cultivating a cell,under conditions conducive for production of the dispersin variants; andoptionally, (b) recovering the dispersin variant. In some aspects, thecell is a Terribacillus cell. In another aspect, the cell is aTerribacillus saccharophilus, Terribacillus goriensis or Terribacillussaccharophilus cell.

The present invention also relates to methods of producing a dispersinvariant of the present invention, comprising (a) cultivating arecombinant host cell of the present invention under conditionsconducive for production of the dispersin variant; and optionally, (b)recovering the dispersin variant.

The host cells are cultivated in a nutrient medium suitable forproduction of the polypeptide using methods known in the art. Forexample, the cells may be cultivated by shake flask cultivation, orsmall-scale or large-scale fermentation (including continuous, batch,fed-batch, or solid state fermentations) in laboratory or industrialfermenters in a suitable medium and under conditions allowing thepolypeptide to be expressed and/or isolated. The cultivation takes placein a suitable nutrient medium comprising carbon and nitrogen sources andinorganic salts, using procedures known in the art. Suitable media areavailable from commercial suppliers or may be prepared according topublished compositions (e.g., in catalogues of the American Type CultureCollection). If the polypeptide is secreted into the nutrient medium,the polypeptide can be recovered directly from the medium. If thepolypeptide is not secreted, it can be recovered from cell lysates.

The dispersin variant may be detected using methods known in the artthat are specific for the dispersin variant polypeptide. These detectionmethods include, but are not limited to, use of specific antibodies,formation of an enzyme product, or disappearance of an enzyme substrate.For example, an enzyme assay may be used to determine the activity ofthe polypeptide.

The dispersin variant polypeptide may be recovered using methods knownin the art. For example, the dispersin variant polypeptide may berecovered from the nutrient medium by conventional procedures including,but not limited to, collection, centrifugation, filtration, extraction,spray-drying, evaporation, or precipitation. In some aspects, afermentation broth comprising the dispersin variant is recovered.

The polypeptide may be purified by a variety of procedures known in theart including, but not limited to, chromatography (e.g., ion exchange,affinity, hydrophobic, chromatofocusing, and size exclusion),electrophoretic procedures (e.g., preparative isoelectric focusing),differential solubility (e.g., ammonium sulfate precipitation),SDS-PAGE, or extraction (see, e.g., Protein Purification, Janson andRyden, editors, VCH Publishers, New York, 1989) to obtain substantiallypure polypeptides.

In an alternative aspect, the polypeptide is not recovered, but rather ahost cell of the present invention expressing the polypeptide is used asa source of the variant.

Compositions

The present invention further relates to a cleaning e.g. detergentcomposition comprising at least one dispersin variant according to theinvention and preferably a detergent adjunct ingredient. The detergentcomposition may be used for improving deep-cleaning effect, includingbut not limited to deep cleaning of an item, for preventing and/orreducing the stickiness of an item, for pretreating stains on the item,for preventing and/or reducing redeposition of soil during a wash cycle,for preventing and/or reducing adherence of soil to an item, formaintaining or improving the whiteness of an item and for preventingand/or reducing malodor from an item. The dispersin variants of theinvention are useful in powder and liquid detergent.

In some aspects of the invention, the detergent adjunct ingredient isselected from the group consisting of surfactants, builders,flocculating aid, chelating agents, dye transfer inhibitors, enzymes,enzyme stabilizers, enzyme inhibitors, catalytic materials, bleachactivators, hydrogen peroxide, sources of hydrogen peroxide, preformedperacids, polymeric dispersing agents, clay soilremoval/anti-redeposition agents, brighteners, suds suppressors, dyes,perfumes, structure elasticizing agents, fabric softeners, carriers,hydrotropes, builders and co-builders, fabric hueing agents,anti-foaming agents, dispersants, processing aids, and/or pigments.

The detergent adjunct ingredient may be a surfactant. One advantage ofincluding a surfactant in a detergent composition comprising a dispersinvariant is that the wash performance is improved. In some aspects, thedetergent adjunct ingredient is a builder or a clay soilremoval/anti-redeposition agent.

In some aspects, detergent adjunct ingredient is an enzyme. Thedetergent composition may comprise one or more enzymes, as specifiedbelow. The one or more enzymes may be selected from the group consistingof proteases, amylases, lipases, cutinases, cellulases, endoglucanases,xyloglucanases, pectinases, pectin lyases, xanthanases, peroxidaes,haloperoxygenases, catalases, nucleases e.g. DNases and RNases andmannanases. Specific enzymes suitable for the detergent compositions ofthe invention are described below.

The cleaning composition may be formulated as a bar, a homogenoustablet, and a tablet having two or more layers, a pouch having one ormore compartments, a regular or compact powder, a granule, a paste, agel, or a regular, compact or concentrated liquid. The detergentcomposition can be a liquid detergent, a powder detergent or a granuledetergent.

Some aspects of the present invention relate to laundry or cleaningcompositions comprising a dispersin, preferably at a level of from about0.000001 wt % to about 1 wt %, from about 0.0001 wt % to about 1 wt %,from about 0.0002 wt % to about 1 wt %, from about 0.0005 wt % to about1 wt %, from about 0.001 wt % to about 1 wt %, from about 0.002 wt % toabout 1 wt %, from about 0.005 wt % to about 1 wt %, preferably fromabout 0.01 wt % to about 0.5 wt %, preferably from 0.0002 wt % to about1 wt % by weight (wt %) of the composition. The amounts are wt % perunit active enzyme e.g. from about 0.00001 wt % to about 1 wt % ofdispersin by weight of the composition.

The concentration of the active enzyme having dispersin activity ispreferably at least 0.00001%, preferably at least 0.00002%, preferablyat least 0.0001 wt %, preferably at least 0.0002 wt %, preferably atleast 0.001 wt %, preferably at least 0.002 wt %, preferably at least0.005 wt %, preferably at least 0.01 wt %, preferably at least 0.02 wt%, preferably at least 0.05 wt % preferably at least 0.1 wt % of thetotal detergent concentration.

The amount enzyme may also be in ppm (mg/L) active enzyme protein. Thus,in one aspect the amount of dispersin in the composition is at least0.00001 ppm, 0.00002 ppm, 0.00005 ppm, 0.0001 ppm, 0.0002 ppm, 0.0005ppm, 0.001 ppm, 0.002 ppm, 0.005 ppm, 0.01 ppm, 0.02 ppm, 0.05 ppm, 0.1ppm, 0.2 ppm, 0.5 ppm, 1 ppm, 2 ppm, 5 ppm, 10 ppm or at least 20 ppmdispersin enzymes. In one aspect, the amount of dispersin in thecomposition is in the range from about 0.00001 ppm to about 10 ppm, orin the range from about 0.0001 ppm to about 2 ppm or in the range fromabout 0.001 ppm to about 2 ppm dispersin enzymes.

In some aspects, the cleaning composition is a liquid or powder laundrydetergent, suitable for e.g. washing at high temperature and/or pH, suchas at or above 40° C. and/or at or above pH 8. In some aspects, thedetergent composition is a liquid or powder laundry detergent, suitablefor e.g. washing at low temperature and/or pH, such as at or below 20°C. and/or pH 6. The cleaning composition may also be formulated as aunit dose detergent and/or compact detergent optionally with minimum orno water. The cleaning composition may also be a dish wash detergent.The laundry and dish wash detergents may be phosphate-free.

Surfactants

A surfactant may be selected among nonionic, anionic and/or amphotericsurfactants as described above, preferably anionic or nonionicsurfactants but also amphoteric surfactants may be used. In general,bleach-stable surfactants are preferred. Preferred anionic surfactantsare sulphate surfactants and in particular alkyl ether sulphates,especially C-9-15 alcohol ethersulfates, C12-15 primary alcoholethoxylate, C8-C16 ester sulphates and C10-C14 ester sulphates, such asmono dodecyl ester sulphates Non-limiting examples of anionicsurfactants include sulfates and sulfonates, in particular, linearalkylbenzenesulfonates (LAS), isomers of LAS, branchedalkylbenzenesulfonates (BABS), phenylalkanesulfonates,alpha-olefinsulfonates (AOS), olefin sulfonates, alkene sulfonates,alkane-2,3-diylbis(sulfates), hydroxyalkanesulfonates and disulfonates,alkyl sulfates (AS) such as sodium dodecyl sulfate (SDS), fatty alcoholsulfates (FAS), primary alcohol sulfates (PAS), alcohol ethersulfates(AES or AEOS or FES, also known as alcohol ethoxysulfates or fattyalcohol ether sulfates), secondary alkanesulfonates (SAS), paraffinsulfonates (PS), ester sulfonates, sulfonated fatty acid glycerolesters, alpha-sulfo fatty acid methyl esters (alpha-SFMe or SES)including methyl ester sulfonate (MES), alkyl- or alkenylsuccinic acid,dodecenyl/tetradecenyl succinic acid (DTSA), fatty acid derivatives ofamino acids, diesters and monoesters of sulfo-succinic acid or salt offatty acids (soap), and combinations thereof.

The anionic surfactants are preferably added to the detergent in theform of salts. Suitable cations in these salts are alkali metal ions,such as sodium, potassium and lithium and ammonium salts, for example(2-hydroxyethyl) ammonium, bis(2-hydroxyethyl) ammonium andtris(2-hydroxyethyl) ammonium salts. Non-limiting examples of nonionicsurfactants include alcohol ethoxylates (AE or AEO), alcoholpropoxylates, propoxylated fatty alcohols (PFA), alkoxylated fatty acidalkyl esters, such as ethoxylated and/or propoxylated fatty acid alkylesters, alkylphenol ethoxylates (APE), nonylphenol ethoxylates (NPE),alkylpolyglycosides (APG), alkoxylated amines, fatty acidmonoethanolamides (FAM), fatty acid diethanolamides (FADA), ethoxylatedfatty acid monoethanolamides (EFAM), propoxylated fatty acidmonoethanolamides (PFAM), polyhydroxyalkyl fatty acid amides, or N-acylN-alkyl derivatives of glucosamine (glucamides, GA, or fatty acidglucamides, FAGA), as well as products available under the trade namesSPAN and TWEEN, and combinations thereof. Commercially availablenonionic surfactants include Plurafac™, Lutensol™ and Pluronic™ fromBASF, Dehypon™ series from Cognis and Genapol™ series from Clariant.

Builders

A builder is preferably selected among phosphates, sodium citratebuilders, sodium carbonate, sodium silicate, sodium aluminosilicate(zeolite). Suitable builders are alkali metal or ammonium phosphates,polyphosphates, phosphonates, polyphosphates, carbonates, bicarbonates,borates, citrates, and polycarboxylates. Citrate builders, e.g., citricacid and soluble salts thereof (particularly sodium salt), arepolycarboxylate builders. Citrates can be used in combination withzeolite, silicates like the BRITESIL types, and/or layered silicatebuilders. The builder is preferably added in an amount of about 0-65% byweight, such as about 5% to about 50% by weight. In a laundry detergent,the level of builder is typically about 40-65% by weight, particularlyabout 50-65% by weight, particularly from 20% to 50% by weight. Thebuilder and/or co-builder may particularly be a chelating agent thatforms water-soluble complexes with Ca and Mg. Any builder and/orco-builder known in the art for use in cleaning detergents may beutilized. Non-limiting examples of builders include zeolites,diphosphates (pyrophosphates), triphosphates such as sodium triphosphate(STP or STPP), carbonates such as sodium carbonate, soluble silicatessuch as sodium metasilicate, layered silicates (e.g., SKS-6 fromHoechst), and (carboxymethyl)inulin (CMI), and combinations thereof.Further non-limiting examples of builders include citrate, chelatorssuch as aminocarboxylates, aminopolycarboxylates and phosphonates, andalkyl- or alkenylsuccinic acid. Additional specific examples include2,2′,2″-nitrilotriacetic acid (NTA), ethylenediaminetetraacetic acid(EDTA), diethylenetriaminepentaacetic acid (DTPA), iminodisuccinic acid(IDS), ethylenediamine-N,N′-disuccinic acid (EDDS),methylglycine-N,N-diacetic acid (MGDA), glutamic acid-N,N-diacetic acid(GLDA), 1-hydroxyethane-1,1-diphosphonic acid,N-(2-hydroxyethyl)iminodiacetic acid (EDG), aspartic acid-N-monoaceticacid (ASMA), aspartic acid-N,N-diacetic acid (ASDA), asparticacid-N-monopropionic acid (ASMP), iminodisuccinic acid (IDA),N-(sulfomethyl)aspartic acid (SMAS), N-(2-sulfoethyl)-aspartic acid(SEAS), N-(sulfomethylglutamic acid (SMGL), N-(2-sulfoethyl)-glutamicacid (SEGL), N-methyliminodiacetic acid (MIDA), serine-N,N-diacetic acid(SEDA), isoserine-N,N-diacetic acid (ISDA), phenylalanine-N,N-diaceticacid (PHDA), anthranilic acid-N,N-diacetic acid (ANDA), sulfanilicacid-N,N-diacetic acid (SLDA) , taurine-N,N-diacetic acid (TUDA) andN′-(2-hydroxyethypethylenediamine-N,N,N′-triacetic acid (HEDTA),diethanolglycine (DEG), and combinations and salts thereof. Phosphonatessuitable for use herein include 1-hydroxyethane-1,1-diphosphonic acid(HEDP), ethylenediaminetetrakis (methylenephosphonicacid) (EDTMPA),diethylenetriaminepentakis (methylenephosphonic acid) (DTMPA or DTPMPAor DTPMP), nitrilotris (methylenephosphonic acid) (ATMP or NTMP),2-phosphonobutane-1,2,4-tricarboxylic acid (PBTC),hexamethylenediaminetetrakis (methylenephosphonic acid) (HDTMP). Thecomposition may also contain 0-50% by weight, such as about 5% to about30%, of a detergent co-builder. The detergent composition may include aco-builder alone, or in combination with a builder, for example azeolite builder. Non-limiting examples of co-builders includehomopolymers of polyacrylates or copolymers thereof, such as poly(acrylic acid) (PAA) or copoly (acrylic acid/maleic acid) (PAA/PMA) orpolyaspartic acid. Further exemplary builders and/or co-builders aredescribed in, e.g., WO 09/102854, US 5977053. In some aspects, thebuilder is a non-phosphorus based builder such as citric acid and/ormethylglycine-N, N-diacetic acid (MGDA) and/or glutamic-N, N-diaceticacid (GLDA) and/or salts thereof. The liquid composition may also bephosphate free in that instance the preferred builders includes citrateand/or methylglycine-N, N-diacetic acid (MGDA) and/or glutamic-N,N-diacetic acid (GLDA) and/or salts thereof.

Bleach Components

The detergent may contain 0-30% by weight, such as about 1% to about20%, of a bleaching system. Any bleaching system comprising componentsknown in the art for use in cleaning detergents may be utilized.Suitable bleaching system components include sources of hydrogenperoxide; sources of peracids; and bleach catalysts or boosters.

Sources of hydrogen peroxide: Suitable sources of hydrogen peroxide areinorganic persalts, including alkali metal salts such as sodiumpercarbonate and sodium perborates (usually mono- or tetrahydrate), andhydrogen peroxide—urea (1/1).

Sources of peracids: Peracids may be (a) incorporated directly aspreformed peracids or (b) formed in situ in the wash liquor fromhydrogen peroxide and a bleach activator (perhydrolysis) or (c) formedin situ in the wash liquor from hydrogen peroxide and a perhydrolase anda suitable substrate for the latter, e.g., an ester.

-   -   a) Suitable preformed peracids include, but are not limited to,        peroxycarboxylic acids such as peroxybenzoic acid and its        ring-substituted derivatives, peroxy-a-naphthoic acid,        peroxyphthalic acid, peroxylauric acid, peroxystearic acid,        c-phthalimidoperoxycaproic acid [phthalimidoperoxyhexanoic acid        (PAP)], and o-carboxybenzamidoperoxycaproic acid; aliphatic and        aromatic diperoxydicarboxylic acids such as        diperoxydodecanedioic acid, diperoxyazelaic acid,        diperoxysebacic acid, diperoxybrassylic acid,        2-decyldiperoxybutanedioic acid, and diperoxyphthalic,        -isophthalic and -terephthalic acids; perimidic acids;        peroxymonosulfuric acid; peroxydisulfuric acid; peroxyphosphoric        acid; peroxysilicic acid; and mixtures of said compounds. It is        understood that the peracids mentioned may in some cases be best        added as suitable salts, such as alkali metal salts (e.g.,        Oxone®) or alkaline earth-metal salts.    -   b) Suitable bleach activators include those belonging to the        class of esters, amides, imides, nitriles or anhydrides and,        where applicable, salts thereof. Suitable examples are        tetraacetylethylenediamine (TAED), sodium        4-[(3,5,5-trimethylhexanoyDoxy]benzene-1-sulfonate (ISONOBS),        sodium 4-(dodecanoyloxy)benzene-1-sulfonate (LOBS), sodium        4-(decanoyloxy)benzene-1 -sulfonate, 4-(decanoyloxy)benzoic acid        (DOBA), sodium 4-(nonanoyloxy)benzene-1 -sulfonate (NOBS),        and/or those disclosed in WO98/17767. A particular family of        bleach activators of interest was disclosed in EP624154 and        particularly preferred in that family is acetyl triethyl citrate        (ATC). ATC or a short chain triglyceride like triacetin has the        advantage that they are environmentally friendly. Furthermore,        acetyl triethyl citrate and triacetin have good hydrolytical        stability in the product upon storage and are efficient bleach        activators. Finally, ATC is multifunctional, as the citrate        released in the perhydrolysis reaction may function as a        builder.

Bleach catalysts and boosters: The bleaching system may also include ableach catalyst or booster. Some non-limiting examples of bleachcatalysts that may be used in the compositions of the present inventioninclude manganese oxalate, manganese acetate, manganese-collagen,cobalt-amine catalysts and manganese triazacyclononane (MnTACN)catalysts; particularly preferred are complexes of manganese with1,4,7-trimethyl-1,4,7-triazacyclononane (Me3-TACN) or1,2,4,7-tetramethyl-1,4,7-triazacyclononane (Me4-TACN), in particularMe3-TACN, such as the dinuclear manganese complex[(Me3-TACN)Mn(O)3Mn(Me3-TACN)](PF6)2, and[2,2′,2″-nitrilotris(ethane-1,2-diylazanylylidene-κN-methanylylidene)triphenolato-κ3O]manganese(III).The bleach catalysts may also be other metal compounds, such as iron orcobalt complexes.

In some aspects, where a source of a peracid is included, an organicbleach catalyst or bleach booster may be used having one of thefollowing formulae:

-   -   (iii) and mixtures thereof; wherein each R1 is independently a        branched alkyl group containing from 9 to 24 carbons or linear        alkyl group containing from 11 to 24 carbons, preferably each R1        is independently a branched alkyl group containing from 9 to 18        carbons or linear alkyl group containing from 11 to 18 carbons,        more preferably each R1 is independently selected from the group        consisting of 2-propylheptyl, 2-butyloctyl, 2-pentylnonyl,        2-hexyldecyl, dodecyl, tetradecyl, hexadecyl, octadecyl,        isononyl, isodecyl, isotridecyl and isopentadecyl.

Other exemplary bleaching systems are described, e.g. in WO 2007/087258,WO 2007/087244, WO 2007/087259, EP 1867708 (Vitamin K) and WO2007/087242. Suitable photobleaches may for example be sulfonated zincor aluminium phthalocyanines.

The choice of detergent components may include, for textile care, theconsideration of the type of textile to be cleaned, the type and/ordegree of soiling, the temperature at which cleaning is to take place,and the formulation of the detergent product. Although componentsmentioned below are categorized by general header according to afunctionality, this is not to be construed as a limitation, as acomponent may comprise additional functionalities as will be appreciatedby the skilled artisan, including the exemplary non-limiting componentsshown in below.

Hydrotropes

The detergent composition may contain 0-10% by weight, for example 0-5%by weight, such as about 0.5 to about 5%, or about 3% to about 5%, of ahydrotrope. Any hydrotrope known in the art for use in detergents may beutilized. Non-limiting examples of hydrotropes include sodiumbenzenesulfonate, sodium p-toluene sulfonate (STS), sodium xylenesulfonate (SXS), sodium cumene sulfonate (SCS), sodium cymene sulfonate,amine oxides, alcohols and polyglycolethers, sodium hydroxynaphthoate,sodium hydroxynaphthalene sulfonate, sodium ethylhexyl sulfate, andcombinations thereof.

Polymers

The detergent composition may contain 0-10% by weight, such as 0.5-5%,2-5%, 0.5-2% or 0.2-1% of a polymer. Any polymer known in the art foruse in detergents may be utilized. The polymer may function as aco-builder as mentioned above, or may provide antiredeposition, fibreprotection, soil release, dye transfer inhibition, grease cleaningand/or anti-foaming properties. Some polymers may have more than one ofthe above-mentioned properties and/or more than one of thebelow-mentioned motifs. Exemplary polymers include(carboxymethyl)cellulose (CMC), poly(vinyl alcohol) (PVA),poly(vinylpyrrolidone) (PVP), poly(ethyleneglycol) or poly(ethyleneoxide) (PEG), ethoxylated poly(ethyleneimine), carboxymethyl inulin(CMI), and polycarboxylates such as PAA, PAA/PMA, poly-aspartic acid,and lauryl methacrylate/acrylic acid copolymers , hydrophobicallymodified CMC (HM-CMC) and silicones, copolymers of terephthalic acid andoligomeric glycols, copolymers of poly(ethylene terephthalate) andpoly(oxyethene terephthalate) (PET-POET), PVP, poly(vinylimidazole)(PVI), poly(vinylpyridine-N-oxide) (PVPO or PVPNO) andpolyvinylpyrrolidone-vinylimidazole (PVPVI). Further exemplary polymersinclude sulfonated polycarboxylates, polyethylene oxide andpolypropylene oxide (PEO-PPO) and diquaternium ethoxy sulfate. Otherexemplary polymers are disclosed in, e.g., WO 2006/130575. Salts of theabove-mentioned polymers are also contemplated.

Fabric Hueing Agents

The detergent composition of the present invention may also includefabric hueing agents such as dyes or pigments, which when formulated indetergent compositions can deposit onto a fabric when said fabric iscontacted with a wash liquor comprising said detergent compositions andthus altering the tint of said fabric through absorption/reflection ofvisible light. Fluorescent whitening agents emit at least some visiblelight. In contrast, fabric hueing agents alter the tint of a surface asthey absorb at least a portion of the visible light spectrum. Suitablefabric hueing agents include dyes and dye-clay conjugates and may alsoinclude pigments. Suitable dyes include small molecule dyes andpolymeric dyes. Suitable small molecule dyes include small molecule dyesselected from the group consisting of dyes falling into the Colour Index(C.I.) classifications of Direct Blue, Direct Red, Direct Violet, AcidBlue, Acid Red, Acid Violet, Basic Blue, Basic Violet and Basic Red, ormixtures thereof, for example as described in WO 2005/03274, WO2005/03275, WO 2005/03276 and EP 1876226 (hereby incorporated byreference). The detergent composition preferably comprises from about0.00003 wt % to about 0.2 wt %, from about 0.00008 wt % to about 0.05 wt%, or even from about 0.0001 wt % to about 0.04 wt % fabric hueingagent. The composition may comprise from 0.0001 wt % to 0.2 wt % fabrichueing agent, this may be especially preferred when the composition isin the form of a unit dose pouch. Suitable hueing agents are alsodisclosed in, e.g. WO 2007/087257 and WO 2007/087243.

Enzymes

The detergent composition may comprise one or more additional enzymessuch as a protease, lipase, cutinase, amylase, carbohydrase, cellulase,pectinase, mannanase, arabinase, galactanase, xylanase, oxidase, e.g., alaccase, and/or peroxidase.

In general, the properties of the selected enzyme(s) should becompatible with the selected detergent, (i.e., pH-optimum, compatibilitywith other enzymatic and non-enzymatic ingredients, etc.), and theenzyme(s) should be present in effective amounts.

Cellulases

Suitable cellulases include those of bacterial or fungal origin.Chemically modified or protein engineered mutants are included. Suitablecellulases include cellulases from the genera Bacillus, Pseudomonas,Humicola, Fusarium, Thielavia, Acremonium, e.g., the fungal cellulasesproduced from Humicola insolens, Myceliophthora thermophila and Fusariumoxysporum disclosed in US 4,435,307, US 5,648,263, US 5,691,178, US5,776,757 and WO 89/09259.

Especially suitable cellulases are the alkaline or neutral cellulaseshaving colour care benefits. Examples of such cellulases are cellulasesdescribed in EP 0 495 257, EP 0 531 372, WO 96/11262, WO 96/29397, WO98/08940. Other examples are cellulase variants such as those describedin WO 94/07998, EP 0 531 315, US 5,457,046, US 5,686,593, US 5,763,254,WO 95/24471, WO 98/12307 and WO 99/001544.

Other cellulases are endo-beta-1,4-glucanase enzyme having a sequence ofat least 97% identity to the amino acid sequence of position 1 toposition 773 of SEQ ID NO: 2 of WO 2002/099091 or a family 44xyloglucanase, which a xyloglucanase enzyme having a sequence of atleast 60% identity to positions 40-559 of SEQ ID NO: 2 of WO2001/062903.

Commercially available cellulases include Celluzyme™, and Carezyme™(Novozymes A/S) Carezyme Premium™ (Novozymes A/S), Celluclean™(Novozymes A/S), Celluclean Classic™ (Novozymes A/S),Cellusoft™(Novozymes A/S), Whitezyme™ (Novozymes A/S), Clazinase™, andPuradax HA™ (Genencor International Inc.), and KAC-500(B)™ (KaoCorporation).

Proteases

Suitable proteases include those of bacterial, fungal, plant, viral oranimal origin e.g. vegetable or microbial origin. Microbial origin ispreferred. Chemically modified or protein engineered mutants areincluded. It may be an alkaline protease, such as a serine protease or aneutral protease such as a metalloprotease. A serine protease may forexample be of the S1 family, such as trypsin, or the S8 family such assubtilisin. A metalloproteases protease may for example be a thermolysinfrom e.g. family M4 or other metalloprotease such as those from M5, M7or M8 families.

The term “subtilases” refers to a sub-group of serine protease accordingto Siezen et al., Protein Engng. 4 (1991) 719-737 and Siezen et al.Protein Science 6 (1997) 501-523. Serine proteases are a subgroup ofproteases characterized by having a serine in the active site, whichforms a covalent adduct with the substrate. The subtilases may bedivided into 6 sub-divisions, i.e. the Subtilisin family, the Thermitasefamily, the Proteinase K family, the Lantibiotic peptidase family, theKexin family and the Pyrolysin family.

Examples of subtilases are those derived from Bacillus such as Bacilluslentus, B. alkalophilus, B. subtilis, B. amyloliquefaciens, Bacilluspumilus and Bacillus gibsonii described in; US7262042 and WO09/021867,and subtilisin lentus, subtilisin Novo, subtilisin Carlsberg, Bacilluslicheniformis, subtilisin BPN', subtilisin 309, subtilisin 147 andsubtilisin 168 described in WO89/06279 and protease PD138 described in(WO93/18140). Other useful proteases may be those described inWO92/175177, WO01/016285, WO02/026024 and WO02/016547. Examples oftrypsin-like proteases are trypsin (e.g. of porcine or bovine origin)and the Fusarium protease described in WO89/06270, WO94/25583 andWO05/040372, and the chymotrypsin proteases derived from Cellumonasdescribed in WO05/052161 and WO05/052146.

A further preferred protease is the alkaline protease from Bacilluslentus DSM 5483, as described for example in WO95/23221, and variantsthereof which are described in WO92/21760, WO95/23221, EP1921147 andEP1921148.

Examples of metalloproteases are the neutral metalloprotease asdescribed in WO07/044993 (Genencor Int.) such as those derived fromBacillus amyloliquefaciens.

Examples of useful proteases are the variants described in: WO92/19729,WO96/034946, WO98/20115, WO98/20116, WO99/011768, WO01/44452,WO03/006602, WO04/03186, WO04/041979, WO07/006305, WO11/036263,WO11/036264, especially the variants with substitutions in one or moreof the following positions: 3, 4, 9, 15, 24, 27, 42, 55, 59, 60, 66, 74,85, 96, 97, 98, 99, 100, 101, 102, 104, 116, 118, 121, 126, 127, 128,154, 156, 157, 158, 161, 164, 176, 179, 182, 185, 188, 189, 193, 198,199, 200, 203, 206, 211, 212, 216, 218, 226, 229, 230, 239, 246, 255,256, 268 and 269 wherein the positions correspond to the positions ofthe Bacillus lentus protease shown in SEQ ID NO: 1 of WO 2016/001449.More preferred the protease variants may comprise one or more of thefollowing mutations: S3T, V4I, S9R, S9E, A15T, S24G, S24R, K27R, N42R,S55P, G59E, G59D, N60D, N60E, V66A, N74D, S85R, A96S, S97G, S97D, S97A,S97SD, S99E, S99D, S99G, S99M, S99N, S99R, S99H, S101A, V102I, V102Y,V102N, S104A, G116V, G116R, H118D, H118N, A120S, S126L, P127Q, S128A,S154D, A156E, G157D, G157P, S158E, Y161A, R164S, Q176E, N179E, S182E,Q185N, A188P, G189E, V193M, N198D, V199I, Y203W, S206G, L211Q, L211D,N212D, N212S, M216S, A226V, K229L, Q230H, Q239R, N246K, N255W, N255D,N255E, L256E, L256D T268A and R269H. The protease variants arepreferably variants of the Bacillus lentus protease (Savinase®) shown inSEQ ID NO: 1 of WO 2016/001449 or the Bacillus amylolichenifaciensprotease (BPN') shown in SEQ ID NO: 2 of WO2016/001449. The proteasevariants preferably have at least 80% sequence identity to thepolypeptide shown in SEQ ID NO: 1 or SEQ ID NO: 2 of WO 2016/001449.

A protease variant comprising a substitution at one or more positionscorresponding to positions 171, 173, 175, 179, or 180 of SEQ ID NO: 1 ofWO2004/067737, wherein said protease variant has a sequence identity ofat least 75% but less than 100% to SEQ ID NO: 1 of WO2004/067737.

Suitable commercially available protease enzymes include those soldunder the trade names Alcalase®, Duralase™, Durazym™, Relase®, Relase®Ultra, Savinase®, Savinase® Ultra, Primase®, Polarzyme®, Kannase®,Liquanase®, Liquanase® Ultra, Ovozyme®, Coronase®, Coronase® Ultra,Blaze®, Blaze Evity® 100T, Blaze Evity® 125T, Blaze Evity® 150T,Neutrase®, Everlase® and Esperase® (Novozymes A/S), those sold under thetradename Maxatase®, Maxacal®, Maxapem®, Purafect Ox®, Purafect OxP®,Puramax®, FN2®, FN3®, FN4®, Excellase®, Excellenz P1000™, ExcellenzP1250™, Eraser®, Preferenz P100™, Purafect Prime®, Preferenz P110™,Effectenz P1000™, Purafect®™, Effectenz P1050™, Purafect Ox®™, EffectenzP2000™, Purafast®, Properase®, Opticlean® and Optimase®(Danisco/DuPont), Axapem™ (Gist-Brocases N.V.), BLAP (sequence shown inFIG. 29 of US5352604) and variants hereof (Henkel AG) and KAP (Bacillusalkalophilus subtilisin) from Kao.

Lipases and Cutinases

Suitable lipases and cutinases include those of bacterial or fungalorigin. Chemically modified or protein engineered mutant enzymes areincluded. Examples include lipase from Thermomyces, e.g. from T.lanuginosus (previously named Humicola lanuginosa) as described inEP258068 and EP305216, cutinase from Humicola, e.g. H. insolens(WO96/13580), lipase from strains of Pseudomonas (some of these nowrenamed to Burkholderia), e.g. P. alcaligenes or P. pseudoalcaligenes(EP218272), P. cepacia (EP331376), P. sp. strain SD705 (WO95/06720 &WO96/27002), P. wisconsinensis (WO96/12012), GDSL-type Streptomyceslipases (WO10/065455), cutinase from Magnaporthe grisea (WO10/107560),cutinase from Pseudomonas mendocina (US5,389,536), lipase fromThermobifida fusca (WO11/084412), Geobacillus stearothermophilus lipase(WO11/084417), lipase from Bacillus subtilis (WO11/084599), and lipasefrom Streptomyces griseus (WO11/150157) and S. pristinaespiralis(WO12/137147).

Other examples are lipase variants such as those described in EP407225,WO92/05249, WO94/01541, WO94/25578, WO95/14783, WO95/30744, WO95/35381,WO95/22615, WO96/00292, WO97/04079, WO97/07202, WO00/34450, WO00/60063,WO01/92502, WO07/87508 and WO09/109500.

Preferred commercial lipase products include Lipolase™, Lipex™; Lipolex™and Lipoclean™ (Novozymes A/S), Lumafast (originally from Genencor) andLipomax (originally from Gist-Brocades).

Still other examples are lipases sometimes referred to asacyltransferases or perhydrolases, e.g. acyltransferases with homologyto Candida antarctica lipase A (WO10/111143), acyltransferase fromMycobacterium smegmatis (WO05/56782), perhydrolases from the CE 7 family(WO09/67279), and variants of the M. smegmatis perhydrolase inparticular the S54V variant used in the commercial product Gentle PowerBleach from Huntsman Textile Effects Pte Ltd (WO10/100028).

Amylases

Suitable amylases which can be used together with the dispersins of theinvention may be an alpha-amylase or a glucoamylase and may be ofbacterial or fungal origin. Chemically modified or protein engineeredmutants are included. Amylases include, for example, alpha-amylasesobtained from Bacillus, e.g., a special strain of Bacilluslicheniformis, described in more detail in GB 1,296,839.

Suitable amylases include amylases having SEQ ID NO: 2 in WO 95/10603 orvariants having 90% sequence identity to SEQ ID NO: 1 thereof. Preferredvariants are described in WO 94/02597, WO 94/18314, WO 97/43424 and SEQID NO: 4 of WO 99/019467, such as variants with substitutions in one ormore of the following positions: 15, 23, 105, 106, 124, 128, 133, 154,156, 178, 179, 181, 188, 190, 197, 201, 202, 207, 208, 209, 211, 243,264, 304, 305, 391, 408, and 444.

Different suitable amylases include amylases having SEQ ID NO: 6 in WO02/010355 or variants thereof having 90% sequence identity to SEQ ID NO:6. Preferred variants of SEQ ID NO: 6 are those having a deletion inpositions 181 and 182 and a substitution in position 193.

Other amylases which are suitable are hybrid alpha-amylase comprisingresidues 1-33 of the alpha-amylase obtained from B.amyloliquefaciensshown in SEQ ID NO: 6 of WO 2006/066594 and residues36-483 of the B. licheniformis alpha-amylase shown in SEQ ID NO: 4 of WO2006/066594 or variants having 90% sequence identity thereof. Preferredvariants of this hybrid alpha-amylase are those having a substitution, adeletion or an insertion in one of more of the following positions: G48,T49, G107, H156, A181, N190, M197, I201, A209 and Q264. Most preferredvariants of the hybrid alpha-amylase comprising residues 1-33 of thealpha-amylase obtained from B. amyloliquefaciens shown in SEQ ID NO: 6of WO 2006/066594 and residues 36-483 of SEQ ID NO: 4 are those havingthe substitutions:

-   -   M 197T;    -   H156Y+A181T+N190F+A209V+Q264S; or    -   G48A+T49I+G107A+H156Y+A181T+N190F+I201F+A209V+Q264S.

Further amylases which are suitable are amylases having SEQ ID NO: 6 inWO 99/019467 or variants thereof having 90% sequence identity to SEQ IDNO: 6. Preferred variants of SEQ ID NO: 6 are those having asubstitution, a deletion or an insertion in one or more of the followingpositions: R181, G182, H183, G184, N195, 1206, E212, E216 and K269.Particularly preferred amylases are those having deletion in positionsR181 and G182, or positions H183 and G184.

Additional amylases which can be used are those having SEQ ID NO: 1, SEQID NO: 3, SEQ ID NO: 2 or SEQ ID NO: 7 of WO 96/023873 or variantsthereof having 90% sequence identity to SEQ ID NO: 1, SEQ ID NO: 2, SEQID NO: 3 or SEQ ID NO: 7. Preferred variants of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO: 3 or SEQ ID NO: 7 are those having a substitution, adeletion or an insertion in one or more of the following positions: 140,181, 182, 183, 184, 195, 206, 212, 243, 260, 269, 304 and 476, using SEQID NO: 2 of WO 96/023873 for numbering. More preferred variants arethose having a deletion in two positions selected from 181, 182, 183 and184, such as 181 and 182, 182 and 183, or positions 183 and 184. Mostpreferred amylase variants of SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 7are those having a deletion in positions 183 and 184 and a substitutionin one or more of positions 140, 195, 206, 243, 260, 304 and 476.

Other amylases which can be used are amylases having SEQ ID NO: 2 of WO08/153815, SEQ ID NO: 10 in WO 01/66712 or variants thereof having 90%sequence identity to SEQ ID NO: 2 of WO 08/153815 or 90% sequenceidentity to SEQ ID NO: 10 in WO 01/66712. Preferred variants of SEQ IDNO: 10 in WO 01/66712 are those having a substitution, a deletion or aninsertion in one of more of the following positions: 176, 177, 178, 179,190, 201, 207, 211 and 264.

Further suitable amylases are amylases having SEQ ID NO: 2 of WO09/061380 or variants having 90% sequence identity to SEQ ID NO: 2thereof. Preferred variants of SEQ ID NO: 2 are those having atruncation of the C-terminus and/or a substitution, a deletion or aninsertion in one of more of the following positions: Q87, Q98, S125,N128, T131, T165, K178, R180, S181, T182, G183, M201, F202, N225, S243,N272, N282, Y305, R309, D319, Q320, Q359, K444 and G475. More preferredvariants of SEQ ID NO: 2 are those having the substitution in one ofmore of the following positions: Q87E,R, Q98R, S125A, N128C, T131I,T165I, K178L, T182G, M201L, F202Y, N225E,R, N272E,R, S243Q,A,E,D, Y305R,R309A, Q320R, Q359E, K444E and G475K and/or deletion in position R180and/or S181 or of T182 and/or G183. Most preferred amylase variants ofSEQ ID NO: 2 are those having the substitutions:

N 128C+K178L+T182G+Y305R+G475K; N128C+K178L+T182G+F202Y+Y305R+D319T+G475K; S125A+N128C+K178L+T182G+Y305R+G475K; or

S125A+N128C+T131I+T1651+K178L+T182G+Y305R+G475K wherein the variants areC-terminally truncated and optionally further comprises a substitutionat position 243 and/or a deletion at position 180 and/or position 181.

Other suitable amylases are the alpha-amylase having SEQ ID NO: 12 in WO01/66712 or a variant having at least 90% sequence identity to SEQ IDNO: 12. Preferred amylase variants are those having a substitution, adeletion or an insertion in one of more of the following positions ofSEQ ID NO: 12 in WO01/66712: R28, R118, N174; R181, G182, D183, G184,G186, W189, N195, M202, Y298, N299, K302, S303, N306, R310, N314; R320,H324, E345, Y396, R400, W439, R444, N445, K446, Q449, R458, N471, N484.Particular preferred amylases include variants having a deletion of D183and G184 and having the substitutions R118K, N195F, R320K and R458K, anda variant additionally having substitutions in one or more positionselected from the group: M9, G149, G182, G186, M202, T257, Y295, N299,M323, E345 and A339, most preferred a variant that additionally hassubstitutions in all these positions.

Further suitable amylases are alpha-amylase variants comprising amodification in one or more positions corresponding to positions 1, 54,56, 72, 109, 113, 116, 134, 140, 159, 167, 169, 172, 173, 174, 181, 182,183, 184, 189, 194, 195, 206, 255, 260, 262, 265, 284, 289, 304, 305,347, 391, 395, 439, 469, 444, 473, 476, or 477 of SEQ ID NO: 1, whereinthe alpha-amylase variant has a sequence identity of at least 75% butless than 100% to SEQ ID NO: 13 or SEQ ID NO: 14 of WO 2016/180748.

Other examples are amylase variants such as those described in WO2011/098531, WO 2013/001078 and WO 2013/001087.

Commercially available amylases are Duramyl™, Termamyl™, Fungamyl™,Stainzyme ™, Stainzyme Plus™, Natalase™, Liquozyme X and BAN™ (fromNovozymes A/S), and Rapidase™, Purastar™/Effectenz™, Powerase andPreferenz S100 (from Genencor International Inc./DuPont).

Peroxidases/Oxidases

A peroxidase according to the invention is a peroxidase enzyme comprisedby the enzyme classification EC 1.11.1.7, as set out by the NomenclatureCommittee of the International Union of Biochemistry and MolecularBiology (IUBMB), or any fragment obtained therefrom, exhibitingperoxidase activity.

Suitable peroxidases include those of plant, bacterial or fungal origin.Chemically modified or protein engineered mutants are included. Examplesof useful peroxidases include peroxidases from Coprinopsis, e.g., fromC. cinerea (EP 179,486), and variants thereof as those described in WO93/24618, WO 95/10602, and WO 98/15257.

A peroxidase according to the invention also includes a haloperoxidaseenzyme, such as chloroperoxidase, bromoperoxidase and compoundsexhibiting chloroperoxidase or bromoperoxidase activity. Haloperoxidasesare classified according to their specificity for halide ions.Chloroperoxidases (E.C. 1.11.1.10) catalyze formation of hypochloritefrom chloride ions.

In an aspect, the haloperoxidase of the invention is a chloroperoxidase.Preferably, the haloperoxidase is a vanadium haloperoxidase, i.e., avanadate-containing haloperoxidase. In a preferred method of the presentinvention the vanadate-containing haloperoxidase is combined with asource of chloride ion.

Haloperoxidases have been isolated from many different fungi, inparticular from the fungus group dematiaceous hyphomycetes, such asCaldariomyces, e.g., C. fumago, Alternaria, Curvularia, e.g., C.verruculosa and C. inaequalis, Drechslera, Ulocladium and Botrytis.

Haloperoxidases have also been isolated from bacteria such asPseudomonas, e.g., P. pyrrocinia and Streptomyces, e.g., S.aureofaciens.

In a preferred aspect, the haloperoxidase is derivable from Curvulariasp., in particular Curvularia verruculosa or Curvularia inaequalis, suchas C. inaequalis CBS 102.42 as described in WO 95/27046; or C.verruculosa CBS 147.63 or C. verruculosa CBS 444.70 as described in WO97/04102; or from Drechslera hartlebii as described in WO 01/79459,Dendtyphiella salina as described in WO 01/79458, Phaeotrichoconiscrotalarie as described in WO 01/79461, or Geniculosporium sp. asdescribed in WO 01/79460.

An oxidase according to the invention include, in particular, anylaccase enzyme comprised by the enzyme classification EC 1.10.3.2, orany fragment obtained therefrom exhibiting laccase activity, or acompound exhibiting a similar activity, such as a catechol oxidase (EC1.10.3.1), an o-aminophenol oxidase (EC 1.10.3.4), ora bilirubin oxidase(EC 1.3.3.5).

Preferred laccase enzymes are enzymes of microbial origin. The enzymesmay be obtained from plants, bacteria or fungi (including filamentousfungi and yeasts).

Suitable examples from fungi include a laccase derivable from a strainof Bacillus, Neurospora, e.g., N. crassa, Podospora, Botrytis, Collybia,Fomes, Lentinus, Pleurotus, Trametes, e.g., T. villosa and T.versicolor, Rhizoctonia, e.g., R. solani, Coprinopsis, e.g., C. cinerea,C. comatus, C. friesii, and C. plicatilis, Psathyrella, e.g., P.condelleana, Panaeolus, e.g., P. papilionaceus, Myceliophthora, e.g., M.thermophila, Schytalidium, e.g., S. thermophilum, Polyporus, e.g., P.pinsitus, Phlebia, e.g., P. radiata (WO 92/01046), or Coriolus, e.g., C.hirsutus (JP 2238885).

Suitable examples from bacteria include a laccase derivable from astrain of Bacillus.

A laccase obtained from Coprinopsis or Myceliophthora is preferred; inparticular a laccase obtained from Coprinopsis cinerea, as disclosed inWO 97/08325; or from Myceliophthora thermophila, as disclosed in WO95/33836.

The detergent enzyme(s) may be included in a detergent composition byadding separate additives containing one or more enzymes, or by adding acombined additive comprising all of these enzymes. A detergent additiveof the invention, i.e., a separate additive or a combined additive, canbe formulated, for example, as a granulate, liquid, slurry, etc.Preferred detergent additive formulations are granulates, in particularnon-dusting granulates, liquids, in particular stabilized liquids, orslurries.

Non-dusting granulates may be produced, e.g. as disclosed in US4,106,991 and 4,661,452 and may optionally be coated by methods known inthe art. Examples of waxy coating materials are poly (ethylene oxide)products (polyethyleneglycol, PEG) with mean molar weights of 1000 to20000; ethoxylated nonylphenols having from 16 to 50 ethylene oxideunits; ethoxylated fatty alcohols in which the alcohol contains from 12to 20 carbon atoms and in which there are 15 to 80 ethylene oxide units;fatty alcohols; fatty acids; and mono- and di- and triglycerides offatty acids. Examples of film-forming coating materials suitable forapplication by fluid bed techniques are given in GB 1483591. Liquidenzyme preparations may, for instance, be stabilized by adding a polyolsuch as propylene glycol, a sugar or sugar alcohol, lactic acid or boricacid according to established methods. Protected enzymes may be preparedaccording to the method disclosed in EP 238,216.

Dispersants

The detergent compositions of the present invention can also containdispersants. In particular powdered detergents may comprise dispersants.Suitable water-soluble organic materials include the homo- orco-polymeric acids or their salts, in which the polycarboxylic acidcomprises at least two carboxyl radicals separated from each other bynot more than two carbon atoms. Suitable dispersants are for exampledescribed in Powdered Detergents, Surfactant science series volume 71,Marcel Dekker, Inc.

Dye Transfer Inhibiting Agents

The cleaning compositions of the present invention may also include oneor more dye transfer inhibiting agents. Suitable polymeric dye transferinhibiting agents include, but are not limited to, polyvinylpyrrolidonepolymers, polyamine N-oxide polymers, copolymers of N-vinylpyrrolidoneand N-vinylimidazole, polyvinyloxazolidones and polyvinylimidazoles ormixtures thereof. When present in a subject composition, the dyetransfer inhibiting agents may be present at levels from about 0.0001%to about 10%, from about 0.01% to about 5% or even from about 0.1% toabout 3% by weight of the composition.

Fluorescent Whitening Agent

The detergent composition may preferably also contain additionalcomponents that may tint articles being cleaned, such as fluorescentwhitening agent or optical brighteners. Where present the brightener ispreferably at a level of about 0.01% to about 0.5%. Any fluorescentwhitening agent suitable for use in a laundry detergent composition maybe used in the composition of the present invention. The most commonlyused fluorescent whitening agents are those belonging to the classes ofdiaminostilbene-sulfonic acid derivatives, diarylpyrazoline derivativesand bisphenyl-distyryl derivatives. Examples of thediaminostilbene-sulfonic acid derivative type of fluorescent whiteningagents include the sodium salts of:4,4′-bis-(2-diethanolamino-4-anilino-s-triazin-6-ylamino)stilbene-2,2′-disulfonate, 4,4′-bis-(2,4-dianilino-s-triazin-6-ylamino)stilbene-2.2′-disulfonate,4,4′-bis-(2-anilino-4-(N-methyl-N-2-hydroxy-ethylamino)-s-triazin-6-ylamino)stilbene-2,2′-disulfonate,4,4′-bis-(4-phenyl-1,2,3-triazol-2-yl)stilbene-2,2′-disulfonate andsodium5-(2H-naphtho[1,2-d][1,2,3]triazol-2-yl)-2-[(E)-2-phenylvinyl]benzenesulfonate.Preferred fluorescent whitening agents are Tinopal DMS and Tinopal CBSavailable from Ciba-Geigy AG, Basel, Switzerland. Tinopal DMS is thedisodium salt of 4,4′-bis-(2-morpholino-4-anilino-s-triazin-6-ylamino)stilbene-2,2′-disulfonate. Tinopal CBS is the disodium salt of2,2′-bis-(phenyl-styryl)-disulfonate. Also preferred are fluorescentwhitening agents is the commercially available Parawhite KX, supplied byParamount Minerals and Chemicals, Mumbai, India. Tinopal CBS-X is a4.4′-bis-(sulfostyryl)-biphenyl disodium salt also known as DisodiumDistyrylbiphenyl Disulfonate. Other fluorescers suitable for use in theinvention include the 1-3-diaryl pyrazolines and the7-alkylaminocoumarins.

Suitable fluorescent brightener levels include lower levels of fromabout 0.01, from 0.05, from about 0.1 or even from about 0.2 wt % toupper levels of 0.5 or even 0.75 wt %.

Soil Release Polymers

The detergent compositions may also include one or more soil releasepolymers which aid the removal of soils from fabrics such as cotton andpolyester based fabrics, in particular the removal of hydrophobic soilsfrom polyester based fabrics. The soil release polymers may for examplebe nonionic or anionic terephthalte based polymers, polyvinylcaprolactam and related copolymers, vinyl graft copolymers, polyesterpolyamides see for example Chapter 7 in Powdered Detergents, Surfactantscience series volume 71, Marcel Dekker, Inc. Another type of soilrelease polymers is amphiphilic alkoxylated grease cleaning polymerscomprising a core structure and a plurality of alkoxylate groupsattached to that core structure. The core structure may comprise apolyalkylenimine structure or a polyalkanolamine structure as describedin detail in WO 2009/087523 (hereby incorporated by reference).Furthermore, random graft co-polymers are suitable soil releasepolymers. Suitable graft co-polymers are described in more detail in WO2007/138054, WO 2006/108856 and WO 2006/113314 (hereby incorporated byreference). Other soil release polymers are substituted polysaccharidestructures especially substituted cellulosic structures such as modifiedcellulose derivatives such as those described in EP 1867808 or WO2003/040279 (both are hereby incorporated by reference). Suitablecellulosic polymers include cellulose, cellulose ethers, celluloseesters, cellulose amides and mixtures thereof. Suitable cellulosicpolymers include anionically modified cellulose, nonionically modifiedcellulose, cationically modified cellulose, zwitterionically modifiedcellulose, and mixtures thereof. Suitable cellulosic polymers includemethyl cellulose, carboxy methyl cellulose, ethyl cellulose, hydroxylethyl cellulose, hydroxyl propyl methyl cellulose, ester carboxy methylcellulose, and mixtures thereof.

Anti-Redeposition Agents

The detergent compositions of the present invention may also include oneor more anti-redeposition agents such as carboxymethylcellulose (CMC),polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), polyoxyethyleneand/or polyethyleneglycol (PEG), homopolymers of acrylic acid,copolymers of acrylic acid and maleic acid, and ethoxylatedpolyethyleneimines. The cellulose based polymers described under soilrelease polymers above may also function as anti-redeposition agents.

Rheology Modifiers

The detergent compositions of the present invention may also include oneor more rheology modifiers, structurants or thickeners, as distinct fromviscosity reducing agents. The rheology modifiers are selected from thegroup consisting of non-polymeric crystalline, hydroxy-functionalmaterials, polymeric rheology modifiers which impart shear thinningcharacteristics to the aqueous liquid matrix of a liquid detergentcomposition. The rheology and viscosity of the detergent can be modifiedand adjusted by methods known in the art, for example as shown in EP2169040.

Other suitable adjunct materials include, but are not limited to,anti-shrink agents, anti-wrinkling agents, bactericides, binders,carriers, dyes, enzyme stabilizers, fabric softeners, fillers, foamregulators, hydrotropes, perfumes, pigments, sod suppressors, solvents,and structurants for liquid detergents and/or structure elasticizingagents.

Other Materials

Any detergent components known in the art for use in the cleaningcomposition of the invention may also be utilized. Other optionaldetergent components include anti-corrosion agents, anti-shrink agents,anti-soil redeposition agents, anti-wrinkling agents, bactericides,binders, corrosion inhibitors, disintegrants/disintegration agents,dyes, enzyme stabilizers (including boric acid, borates, CMC, and/orpolyols such as propylene glycol), fabric conditioners including clays,fillers/processing aids, fluorescent whitening agents/opticalbrighteners, foam boosters, foam (suds) regulators, perfumes,soil-suspending agents, softeners, suds suppressors, tarnish inhibitors,and wicking agents, either alone or in combination. Any ingredient knownin the art for use in detergents may be utilized. The choice of suchingredients is well within the skill of the artisan.

Formulation of Detergent Products

The detergent composition may be in any convenient form, e.g., a bar, ahomogenous tablet, a tablet having two or more layers, a regular orcompact powder, a granule, a paste, a gel, or a regular, compact orconcentrated liquid.

Detergent formulation forms: Layers (same or different phases), Pouches,versus forms for Machine dosing unit.

Pouches can be configured as single or multicompartments. It can be ofany form, shape and material which is suitable for hold the composition,e.g. without allowing the release of the composition to release of thecomposition from the pouch prior to water contact. The pouch is madefrom water soluble film which encloses an inner volume. Said innervolume can be divided into compartments of the pouch. Preferred filmsare polymeric materials preferably polymers which are formed into a filmor sheet. Preferred polymers, copolymers or derivates thereof areselected polyacrylates, and water soluble acrylate copolymers, methylcellulose, carboxy methyl cellulose, sodium dextrin, ethyl cellulose,hydroxyethyl cellulose, hydroxypropyl methyl cellulose, malto dextrin,poly methacrylates, most preferably polyvinyl alcohol copolymers and,hydroxyprpyl methyl cellulose (HPMC). Preferably the level of polymer inthe film for example PVA is at least about 60%. Preferred averagemolecular weight will typically be about 20,000 to about 150,000. Filmscan also be of blend compositions comprising hydrolytically degradableand water soluble polymer blends such as polyactide and polyvinylalcohol (known under the Trade reference M8630 as sold by Chris CraftIn. Prod. Of Gary, Ind., US) plus plasticisers like glycerol, ethyleneglycerol, Propylene glycol, sorbitol and mixtures thereof. The pouchescan comprise a solid laundry cleaning composition or part componentsand/or a liquid cleaning composition or part components separated by thewater soluble film. The compartment for liquid components can bedifferent in composition than compartments containing solids. Ref: (US2009/0011970 A1)

Detergent ingredients can be separated physically from each other bycompartments in water dissolvable pouches or in different layers oftablets. Thereby negative storage interaction between components can beavoided. Different dissolution profiles of each of the compartments canalso give rise to delayed dissolution of selected components in the washsolution.

Definition/Characteristics of the Forms:

A liquid or gel detergent, which is not unit dosed, may be aqueous,typically containing at least 20% by weight and up to 95% water, such asup to about 70% water, up to about 65% water, up to about 55% water, upto about 45% water, up to about 35% water. Other types of liquids,including without limitation, alkanols, amines, diols, ethers andpolyols may be included in an aqueous liquid or gel. An aqueous liquidor gel detergent may contain from 0-30% organic solvent.

A liquid or gel detergent may be non-aqueous.

Granular Detergent Formulations

A granular detergent may be formulated as described in WO 09/092699, EP1705241, EP 1382668, WO 07/001262, US 6472364, WO 04/074419 or WO09/102854. Other useful detergent formulations are described in WO09/124162, WO 09/124163, WO 09/117340, WO 09/117341, WO 09/117342, WO09/072069, WO 09/063355, WO 09/132870, WO 09/121757, WO 09/112296, WO09/112298, WO 09/103822, WO 09/087033, WO 09/050026, WO 09/047125, WO09/047126, WO 09/047127, WO 09/047128, WO 09/021784, WO 09/010375, WO09/000605, WO 09/122125, WO 09/095645, WO 09/040544, WO 09/040545, WO09/024780, WO 09/004295, WO 09/004294, WO 09/121725, WO 09/115391, WO09/115392, WO 09/074398, WO 09/074403, WO 09/068501, WO 09/065770, WO09/021813, WO 09/030632, and WO 09/015951.

WO 2011025615, WO 2011016958, WO 2011005803, WO 2011005623, WO2011005730, WO 2011005844, WO 2011005904, WO 2011005630, WO 2011005830,WO 2011005912, WO 2011005905, WO 2011005910, WO 2011005813, WO2010135238, WO 2010120863, WO 2010108002, WO 2010111365, WO 2010108000,WO 2010107635, WO 2010090915, WO 2010033976, WO 2010033746, WO2010033747, WO 2010033897, WO 2010033979, WO 2010030540, WO 2010030541,WO 2010030539, WO 2010024467, WO 2010024469, WO 2010024470, WO2010025161, WO 2010014395, WO 2010044905,

WO 2010145887, WO 2010142503, WO 2010122051, WO 2010102861, WO2010099997, WO 2010084039, WO 2010076292, WO 2010069742, WO 2010069718,WO 2010069957, WO 2010057784, WO 2010054986, WO 2010018043, WO2010003783, WO 2010003792,

WO 2011023716, WO 2010142539, WO 2010118959, WO 2010115813, WO2010105942, WO 2010105961, WO 2010105962, WO 2010094356, WO 2010084203,WO 2010078979, WO 2010072456, WO 2010069905, WO 2010076165, WO2010072603, WO 2010066486, WO 2010066631, WO 2010066632, WO 2010063689,WO 2010060821, WO 2010049187, WO 2010031607, WO 2010000636.

Formulation of Enzyme in Co-Granule

The dispersin may be formulated as a granule for example as a co-granulethat combines one or more enzymes. Each enzyme will then be present inmore granules securing a more uniform distribution of enzymes in thedetergent. This also reduces the physical segregation of differentenzymes due to different particle sizes. Methods for producingmulti-enzyme co-granulate for the detergent industry is disclosed in theIP.com disclosure IPCOM000200739D.

Another example of formulation of enzymes by the use of co-granulatesare disclosed in WO 2013/188331, which relates to a detergentcomposition comprising (a) a multi-enzyme co- granule; (b) less than 10wt zeolite (anhydrous basis); and (c) less than 10 wt phosphate salt(anhydrous basis), wherein said enzyme co-granule comprises from 10 to98 wt % moisture sink components and the composition additionallycomprises from 20 to 80 wt % detergent moisture sink components. WO2013/188331 also relates to a method of treating and/or cleaning asurface, preferably a fabric surface comprising the steps of (i)contacting said surface with the detergent composition as claimed anddescribed herein aqueous wash liquor, (ii) rinsing and/or drying thesurface.

The invention is further described in the following non-limitingparagraphs:

1. A dispersin variant, comprising an alteration, preferablysubstitution at one or more positions corresponding to positions 2, 12,15, 17, 18, 19, 22, 23, 24, 25, 26, 30, 32, 34, 43, 44, 45, 49, 52, 54,56, 57, 58, 59, 60, 62, 63, 67, 68, 71, 72, 74, 77, 79, 80, 81, 82, 90,99, 100, 103, 104, 105, 106, 107, 110, 111, 113, 114, 116, 117, 118,119, 120, 122, 123, 124, 125, 126, 127, 128, 131, 135, 138, 139, 142,145, 147, 148, 149, 150, 151, 152, 163, 164, 167, 168, 170, 171, 173,174, 175, 177, 178, 179, 181, 185, 186, 187, 188, 189, 199, 200, 203,204, 205, 207, 208, 210, 215, 217, 218, 221, 222, 224, 227, 230, 233,234, 235, 237, 244, 249, 251, 252, 253, 254, 256, 260, 261, 262, 263,264, 265, 267, 268, 270, 271, 272, 273, 274, 276, 278, 279, 280, 281,282, 283, 284, 287, 288, 290, 291, 296, 300, 301, 303, 304, 305, 306,308, 309, 312, 314, 315, 319, 321 and 323 of the polypeptide of SEQ IDNO: 1, wherein the dispersin variant has beta-1,6N-acetylglucosaminidase activity, wherein the dispersin variant has atleast 80%, at least 85%, at least 90%, at least 95%, at least 96%, atleast 97%, at least 98%, or at least 99%, but less than 100% sequenceidentity to the polypeptide shown in SEQ ID NO: 1, and wherein thevariant has increased stability, preferably thermo stability measured ashalf-life improvement factor, HIF, compared to the dispersin of SEQ IDNO: 1.2. The variant of paragraph 1, comprising at least one alterationselected from the group consisting of: D2A, D2L, D2N, D2R, D2V, D2W,Q3I, Q3L, Q3M, Q3P, Q3V, Q3Y, S12A, H15F, H15Y, T17C, T17E, T17F, T17M,T17R, T17V, V18L, E19D, E19N, E19P, K22A, K22M, K22V, S23C, S23E, S23I,S23L, S23R, S23T, S23V, V25R, D26M, Y30*, Y30D, Y30L, Y30M, Y30N, Y30R,Y30T, Y30V, G32L, G32M, G32R, N43*, N43H, N43L, E44*, N45D, N45L, N45V,A49W, A49Y, Y52*, Y52M, G54L, G54M, G54N, S56T, S56W, S57W, E58N, N59A,N59C, N59D, N59E, N59F, N59M, N59R, N59V, N59W, T60V, N62C, N62D, N62H,N62Q, N62W, T63C, T63D, T63L, T63N, T63R, T63V, K67A, K67L, N68L, N68Q,L71H, L71N, L71R, L71V, L71W, S72*, S72C, S72D, S72E, S72F, S72G, S72I,S72M, S72N, S72R, S72T, S72Y, I74L, S77A, D79V, K80*, K80E, K80H, K80L,K80N, K80Q, K80V, K80W, D81A, D81G, D81L, D81R, D81S, D81T, D81V, D81W,I82V, L90F, E99Q, E99R, L100S, K103A, K103R, K104N, K104W, D105N, V106A,V106D, V106E, V106H, V106K, V106L, V106M, V106N, V106Q, V106W, V106Y,K107A, K107C, K107L, K107M, K107T, K107V, K107W, N110M, N110R, N110V,D111A, D111E, D111M, D111N, D111Q, D111R, D111V, D111W, V113T, T114C,T114S, Y116D, Y116N, Y116R, S117*, S117D, S117H, S117N, S117P, E118*,E118A, E118D, E118G, E118L, E119G, E119W, T120I, T120L, T120M, T120V,T120W, D122*, D122H, D122R, Y123W, Y124C, Y124I, Y124K, Y124L, Y124M,Y124Q, Y124R, Y124T, Y124V, Y124W, D125C, D125G, D125K, D125Q, D125R,N126V, R127D, R127H, R127K, R127L, R127M, R127Q, R127W, V128C, V128L,V128T, D131V, Q135*, Q135A, Q135D, Q135E, Q135K, Q135M, Q135Y, D138K,D138L, D138M, D138Q, D138R, D138S, D138V, D138W, E139W, D142R, D142W,Y145*, Y145H, Y145L, Y145N, Y145V, P147A, P147C, P147D, P147F, P147G,P147L, P147M, P147R, P147S, P147T, P147V, K148A, K148D, K148L, K148V,F149L, F149M, F149N, E150D, E150H, E150K, E150L, E150M, E150N, E150R,E150V, E150W, E150Y, G151A, G151C, G151D, G151L, G151N, G151P, G151S,G151W, K152D, K152L, K152R, G164D, G164E, G164H, G164S, G164V, V167D,V167E, V167L, V167P, V167Q, V167R, V167W, H168N, L170A, L170D, L170E,L170F, L170H, L170K, L170M, L170N, L170P, L170Q, L170R, L170S, L170V,L170W, L170Y, D171A, D171C, D171E, D171K, D171L, D171M, D171Q, D171R,D171V, D171W, D171Y, I173C, D174H, D174M, D174N, D174V, D174W, F175Y,N177M, Q178*, Q178A, Q178K, Q178R, Q178W, I179T, S181C, S181D, S181F,S181G, S181N, S181P, S181Q, S181T, S181V, S181W, E185M, E185R, E185V,E185W, S186D, S186E, S186H, S186I, S186K, S186L, S186M, S186N, S186Q,S186R, S186V, S186W, K187C, K187D, K187G, K187R, K187S, K187V, K187W,Y188P, E189L, E189V, E189W, S199C, S199L, S199M, S199Y, E200D, E200F,E200K, E200L, E200M, E200N, E200R, E200W, A203C, A203D, A203E, A203G,A203L, A203M, A203P, A203R, A203S, A203T, A203V, A203W, N204L, N204M,N204V, N204W, N204Y, L205I, D207A, D207C, D207E, D207G, D207N, D207Q,D207S, D207V, D207W, S208A, S208C, S208D, S208G, S208L, S208Q, S208T,S208V, S208W, S210T, Q215R, Q215M, Q215L, Q215*, S217V, T218A, T218L,T218Q, T218R, T218V, G222D, E224A, E224P, N227A, N227Q, N227R, N227S,N227T, N227K, D230*, D230R, D230T, D230W, N233H, N233Q, N233R, N233W,W234R, G235A, G235E, G235F, G235H, G235I, G235L, G235M, G235N, G235P,G235S, G235V, S237C, S237G, S237M, S237N, S237W, S237Y, Y244C, Y244E,Y244M, L249H, L249K, L249Q, L249R, L249W, L249Y, S251L, S251N, S251R,S251W, N252P, N252C, G253D, G253W, F254I, F254L, F254M, F254Y, Q256E,Q256R, N260*, N260A, N260C, N260E, N260I, N260K, N260L, N260M, N260Q,N260R, N260V, N260W, N260Y, E261*, E261A, E261D, E261R, E261W, Q262*,Q262F, Q262H, Q262W, Q262Y, M263K, M263L, M263Q, D264*, D264C, D264E,Y265F, N267S, N267T, W268C, W268E, W268M, W268R, Y270F, A271D, A271G,H272D, H272I, H272M, H272P, H272V, N273W, K274R, K274A, K274H, F276A,F276C, F276K, F276N, F276G, F276L, F276M, F276P, F276S, F276V, F276W,I278A, I278K, I278N, I278Q, I278V, S279C, S279D, S279E, S279G, S279N,D280C, D280E, Y281*, Y281A, Y281C, Y281H, Y281K, Y281N, Y281P, Y282E,Y282N, H283I, A284I, A284L, A284N, A284P, A284T, A284V, T287N, S288D,S288K, S288N, V290I, K291L, K291R, K291V, T296C, E300A, E300D, H301C,H301N, H301R, T303A, T303C, T303G, T303K, T303Q, T303R, T303W, D304C,D304M, L305M, L305N, S306C, K308A, K308D, K308G, K308I, K308L, K308Q,K308S, K308T, K308V, K308Y, K309A, K309C, K309D, K309H, K309L, K309M,K309N, K309Q, K309S, K309T, K312A, K312L, K312M, K312N, K312Q, K312S,K312W, E314I, E314L, E314V, L315I, L315V, R319A, and N323R, wherein thepositions correspond to amino acid positions in the amino acid sequenceset forth in SEQ ID NO: 1; and wherein the substitution at the one ormore positions provides a dispersin variant having an increase instability, preferably thermo-stability measured as improvement factor,HIF, of at least 1.1, and further wherein the variants have at least80%, at least 85%, at least 90%, at least 95% identity, at least 96%, atleast 97%, at least 98%, or at least 99%, but less than 100%, sequenceidentity to the polypeptide shown in SEQ ID NO: 1.3. The dispersin variant according to any of the preceding paragraphswherein the variant is obtained from Terribacillus, preferablyTerribacillus saccharophilus, Terribacillus goriensis or Terribacillussaccharophilus.4. The dispersin variant according to any of the preceding paragraphs,wherein the variant comprises one or more motifs: GXDE (SEQ ID NO:8),[EQ][NRSHA][YVFL][AGSTC][IVLF][EAQYN][SN] (SEQ IDNO:9),[VIM][LIV]G[GAV]DE[VI][PSA] SEQ ID NO: 10), [WND[SQR][IVL][TLVM](SEQ ID NO: 11), QSTL (SEQ ID NO: 12), NKFFY (SEQ ID NO: 13) or NLD[DR]S(SEQ ID NO: 14).5. The dispersin variant according to any of the preceding paragraphs,wherein the dispersin variant has improved stability, measured as havingan improvement factor e.g. RAR or HIF>1.0, compared to the dispersinhaving the polypeptide sequence shown in SEQ ID NO: 1 or shown in SEQ IDNO: 15.6. The dispersin variant according to any of the preceding paragraphs,wherein the Half-life Improvement Factor (HIF) is at least 1.2 e.g. 1.5,compared to SEQ ID NO: 1 or compared to SEQ ID NO: 15, wherein HIF iscalculated as T_(1/2dispersinvariant)T_(1/2reference) and the half-life(T_(1/2) in minutes) for the dispersin variants and the dispersinreference (e.g. SEQ ID NO: 1 or 15) is calculated as: 20 minutes ×LN(0.5)/LN(RA) and the residual activity (RA) for each dispersin variantand the reference dispersin (e.g. SEQ ID NO: 1 or 15) is calculated as:slope (stress sample, 56° C. for 60 min)/slope(unstressed sample, 21° C.for 60 min), e.g. as described in Example 2b.7. The dispersin variant of any of the preceding paragraphs, wherein thetotal number of substitutions compared to SEQ ID NO: 1 is 3-20, e.g. atleast 5 to 20, at least 10 to 20 or such as at least 15 to 20substitutions compared to the dispersin comprising the amino acidsequence shown in SEQ ID NO: 1.8. A method for obtaining a dispersin variant, comprising;

-   -   a) introducing into a dispersin parent having 1,6-N-acetyl        glycosaminidase activity, an alteration, preferably a        substitution at one or more positions corresponding to positions        2, 12, 15, 17, 18, 19, 22, 23, 24, 25, 26, 30, 32, 34, 43, 44,        45, 49, 52, 54, 56, 57, 58, 59, 60, 62, 63, 67, 68, 71, 72, 74,        77, 79, 80, 81, 82, 90, 99, 100, 103, 104, 105, 106, 107, 110,        111, 113, 114, 116, 117, 118, 119, 120, 122, 123, 124, 125, 126,        127, 128, 131, 135, 138, 139, 142, 145, 147, 148, 149, 150, 151,        152, 163, 164, 167, 168, 170, 171, 173, 174, 175, 177, 178, 179,        181, 185, 186, 187, 188, 189, 199, 200, 203, 204, 205, 207, 208,        210, 215, 217, 218, 221, 222, 224, 227, 230, 233, 234, 235, 237,        244, 249, 251, 252, 253, 254, 256, 260, 261, 262, 263, 264, 265,        267, 268, 270, 271, 272, 273, 274, 276, 278, 279, 280, 281, 282,        283, 284, 287, 288, 290, 291, 296, 300, 301, 303, 304, 305, 306,        308, 309, 312, 314, 315, 319, 321 and 323 of the polypeptide of        SEQ ID NO: 1,    -   b) and recovering the variant.        9. The method according to paragraph 8, comprising introducing        one or more alteration(s) selected from the group consisting of:        D2A, D2L, D2N, D2R, D2V, D2W, Q3I, Q3L, Q3M, Q3P, Q3V, Q3Y,        S12A, H15F, H15Y, T17C, T17E, T17F, T17M, T17R, T17V, V18L,        E19D, E19K, E19N, E19P, K22A, K22M, K22V, S23A, S23C, S23E,        S23I, S23L, S23R, S23T, S23V, L24I, V25R, D26M, Y30*, Y30A,        Y30D, Y30L, Y30M, Y30N, Y30R, Y30T, Y30V, G32L, G32M, G32R,        N34D, N43*, N43H, N43L, E44*, N45D, N45L, N45V, A49W, A49Y,        Y52*, Y52M, G54L, G54M, G54N, S56T, S56W, S57W, E58N, N59A,        N59C, N59D, N59E, N59F, N59M, N59R, N59V, N59W, T60V, N62C,        N62D, N62H, N62Q, N62W, T63C, T63D, T63L, T63N, T63R, T63V,        K67A, K67L, N68L, N68Q, L71H, L71N, L71R, L71V, L71W, S72*,        S72C, S72D, S72E, S72F, S72G, S72I, S72M, S72N, S72R, S72T,        S72Y, I74L, S77A, D79V, K80*, K80E, K80H, K80L, K80N, K80Q,        K80R, K80V, K80W, D81A, D81G, D81L, D81N, D81R, D81S, D81T,        D81V, D81W, I82V, L90F, E99Q, E99R, L100S, K103A, K103R, K103V,        K104N, K104W, D105N, V106A, V106D, V106E, V106H, V106K, V106L,        V106M, V106N, V106Q, V106R, V106W, V106Y, K107A, K107C, K107L,        K107M, K107T, K107V, K107W, N110M, N110R, N110V, D111A, D111E,        D111M, D111N, D111Q, D111R, D111V, D111W, V113T, T114C, T114S,        Y116D, Y116N, Y116R, S117*, S117D, S117H, S117N, S117P, E118*,        E118A, E118D, E118G, E118L, E119G, E119W, T120I, T120L, T120M,        T120V, T120W, D122*, D122H, D122R, Y123W, Y124C, Y124H, Y124I,        Y124K, Y124L, Y124M, Y124N, Y124Q, Y124R, Y124T, Y124V, Y124W,        D125C, D125G, D125H, D125K, D125Q, D125R, N126V, R127D, R127H,        R127K, R127L, R127M, R127Q, R127W, V128A, V128C, V128D, V128L,        V128T, D131V, Q135*, Q135A, Q135D, Q135E, Q135K, Q135M, Q135Y,        D138K, D138L, D138M, D138N, D138Q, D138R, D138S, D138V, D138W,        E139W, D142R, D142W, Y145*, Y145H, Y145L, Y145N, Y145V, P147A,        P147C, P147D, P147F, P147G, P147L, P147M, P147R, P147S, P147T,        P147V, K148A, K148D, K148L, K148V, F149L, F149M, F149N, E150A,        E150D, E150H, E150K, E150L, E150M, E150N, E150R, E150V, E150W,        E150Y, G151A, G151C, G151D, G151L, G151N, G151P, G151S, G151W,        K152D, K152L, K152R, S163P, G164D, G164E, G164H, G164S, G164V,        V167A, V167D, V167E, V167L, V167P, V167Q, V167R, V167W, H168N,        L170A, L170D, L170E, L170F, L170H, L170K, L170M, L170N, L170P,        L170Q, L170R, L170S, L170V, L170W, L170Y, D171A, D171C, D171E,        D171K, D171L, D171M, D171Q, D171R, D171V, D171W, D171Y, I173C,        D174H, D174M, D174N, D174V, D174W, F175Y, N177M, Q178*, Q178A,        Q178K, Q178R, Q178W, I179T, S181C, S181D, S181F, S181G, S181K,        S181N, S181P, S181Q, S181T, S181V, S181W, E185A, E185M, E185R,        E185V, E185W, S186D, S186E, S186H, S186I, S186K, S186L, S186M,        S186N, S186Q, S186R, S186V, S186W, K187C, K187D, K187G, K187R,        K187S, K187V, K187W, Y188P, E189L, E189V, E189W, S199C, S199L,        S199M, S199Y, E200D, E200F, E200K, E200L, E200M, E200N, E200R,        E200W, A203C, A203D, A203E, A203G, A203L, A203M, A203P, A203Q,        A203R, A203S, A203T, A203V, A203W, N204L, N204M, N204V, N204W,        N204Y, L205I, D207A, D207C, D207E, D207G, D207K, D207N, D207Q,        D207R, D207S, D207V, D207W, S208A, S208C, S208D, S208G, S208L,        S208Q, S208T, S208V, S208W, S210T, Q215K, Q215R, Q215M, Q215L,        Q215*, S217V, T218A, T218L, T218Q, T218R, T218V, S221N, G222D,        E224A, E224P, N227A, N227Q, N227R, N227S, N227T, N227K, D230*,        D230N, D230R, D230T, D230W, N233D, N233E, N233H, N233Q, N233R,        N233W, W234R, G235A, G235E, G235F, G235H, G235I, G235L, G235M,        G235N, G235P, G235S, G235V, S237C, S237G, S237M, S237N, S237W,        S237Y, Y244C, Y244E, Y244M, L249H, L249K, L249Q, L249R, L249W,        L249Y, S251A, S251L, S251N, S251R, S251W, N252P, N252C, G253D,        G253W, F254I, F254L, F254M, F254Y, Q256D, Q256E, Q256R, N260*,        N260A, N260C, N260E, N260I, N260K, N260L, N260M, N260Q, N260R,        N260T, N260V, N260W, N260Y, E261*, E261A, E261D, E261R, E261W,        Q262*, Q262F, Q262H, Q262W, Q262Y, M263K, M263L, M263Q, D264*,        D264C, D264E, D264N, Y265F, N267S, N267T, W268C, W268E, W268M,        W268R, Y270F, A271D, A271G, H272D, H272I, H272M, H272P, H272V,        N273W, K274R, K274A, K274H, F276A, F276C, F276K, F276N, F276G,        F276L, F276M, F276P, F276S, F276V, F276W, I278A, I278K, I278N,        I278Q, I278V, S279C, S279D, S279E, S279G, S279N, D280C, D280E,        Y281*, Y281A, Y281C, Y281H, Y281K, Y281N, Y281P, Y282E, Y282N,        H283I, A284I, A284L, A284N, A284P, A284T, A284V, T287N, S288D,        S288K, S288N, V290I, K291L, K291R, K291V, T296C, E300A, E300D,        H301C, H301N, H301R, T303A, T303C, T303G, T303K, T303Q, T303R,        T303W, D304C, D304M, L305M, L305N, S306C, K308A, K308D, K308E,        K308G, K308I, K308L, K308Q, K308S, K308T, K308V, K308Y, K309A,        K309C, K309D, K309E, K309G, K309H, K309L, K309M, K309N, K309Q,        K309S, K309T, K312A, K312E, K312L, K312M, K312N, K312Q, K312S,        K312W, E314I, E314L, E314V, L315I, L315V, R319A, Y321F and        N323R.        10. The method according to paragraph 8 or 9, wherein the        variant comprises 2-20, e.g. 2-10 and 2-5, such as 2, 3, 4, 5,        6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20        alterations compared to the polypeptide shown in SEQ ID NO: 1.        11. The method according to any of paragraphs 8 to 10, wherein        the dispersin parent obtained from Terribacillus, preferably        Terribacillus saccharophilus, Terribacillus goriensis or        Terribacillus saccharophilus.        12. The method according to any of paragraphs 8 to 11, wherein        the dispersin parent comprises one or more motifs: GXDE (SEQ ID        NO:8), [EQ][NRSHA][YVFL][AGSTC][IVLF][EAQYN][SN] (SEQ ID        NO:9),[VIM][LIV]G[GAV]DE[VI][PSA] SEQ ID NO: 10),        [WND[SQR][IVL][TLVM] (SEQ ID NO: 11), QSTL (SEQ ID NO: 12),        NKFFY (SEQ ID NO: 13) or NLD[DR]S (SEQ ID NO: 14).        13. A method according to any of paragraphs 8 to 12, wherein the        parent dispersin is selected from the group of polypeptides:    -   a) a polypeptide comprising the amino acid sequence shown in SEQ        ID NO: 1 or is a polypeptide having at least 60%, at least 65%,        at least 70%, at least 75%, at least 80%, at least 85%, at least        90%, at least 95%, such as at least 96%, at least 97%, at least        98% or 100% sequence identity hereto,    -   b) a polypeptide comprising the amino acid sequence shown in SEQ        ID NO:2 or is a polypeptide having at least 60%, at least 65%,        at least 70%, at least 75%, at least 80%, at least 85%, at least        90%, at least 95%, such as at least 96%, at least 97%, at least        98% or 100% sequence identity hereto,    -   c) a polypeptide comprising the amino acid sequence shown in SEQ        ID NO:3 or is a polypeptide having at least 95%, such as at        least 96%, such as at least 60%, at least 65%, at least 70%, at        least 75%, at least 80%, at least 85%, at least 90%, at least        95%, such as at least 96%, at least 97%, at least 98% or 100%        sequence identity hereto,    -   d) a polypeptide comprising the amino acid sequence shown in SEQ        ID NO:4 or is a polypeptide having at least 60%, at least 65%,        at least 70%, at least 75%, at least 80%, at least 85%, at least        90%, at least 95%, such as at least 96%, at least 97%, at least        98% or 100% sequence identity hereto, and    -   e) a polypeptide comprising the amino acid sequence shown in SEQ        ID NO:5 or is a polypeptide having at least 60%, at least 65%,        at least 70%, at least 75%, at least 80%, at least 85%, at least        90%, at least 95%, such as at least 96%, at least 97%, at least        98% or 100% sequence identity hereto.        14. A composition, preferably a cleaning composition comprising        a variant according to any of paragraphs 1 to 7.        15. A method of cleaning an item comprising a) adding a        dispersin variant according to any of paragraphs 1 to 7 to a        cleaning composition and b) washing item with the composition,        wherein the item is a textile or a hard surface.        16. A method for finding improved polypeptide variants,        comprising the steps of:    -   a) generate diversity in selected, preferably all, positions of        a polypeptide, thereby preparing libraries of polypeptide        variants (V_(L)), and    -   b) select subset library (V_(LSub)) with improved variants.        17. A method for finding improved polypeptide variants,        comprising the steps of:    -   a) generate diversity in selected, preferably all, positions of        a polypeptide, thereby preparing a library of polypeptide        variants (V_(L)),    -   b) expose part of the library of polypeptide variants (V_(L)) to        one or more stress conditions suitable for measuring a property        of interest, thereby generating a stressed V_(LS+n) and an        unstressed V_(LUS) library;    -   c) compare the stressed V_(LS) and the unstressed V_(LUS)        library; and    -   d) select subset library (V_(LSub)) with improved variants.        18. The method according to any of paragraphs 16 or 17, wherein        the number of variants in the subset library (V_(LSub)) is in        the range of 2-20 polypeptide variants, preferable in the range        of 5-20 polypeptide variants, preferable in the range of 5-10        polypeptide variants, preferable in the range of 8-20        polypeptide variants or in the range of 8-15 polypeptide        variants.        19. The method according to any of paragraphs 16 to 18, wherein        stress condition is preferably elevated temperature such as        above 37° C., above 41° C., above 46° C. or above 50° C.        20. The method according to any of any of paragraphs 16 to 19,        wherein the polypeptide variants are selected from the dispersin        variants of any of paragraphs 1 to 7.

METHODS Assays and Detergent Compositions Composition of Model CetergentA (Liquid)

Ingredients: 12% LAS, 11% AEO Biosoft N25-7 (NI), 7% AEOS (SLES), 6% MPG(monopropylene glycol), 3% ethanol, 3% TEA, 2.75% cocoa soap, 2.75% soyasoap, 2% glycerol, 2% sodium hydroxide, 2% sodium citrate, 1% sodiumformiate, 0.2% DTMPA and 0.2% PCA (all percentages are w/w (weigthpercent %))

Unless otherwise mentioned the tested variants were made from asite-saturation library as described below.

Construction of SSL

A site-saturation library was constructed for each amino acid positionin the enzyme. The resulting transformants were plated onto LB agarplates containing with 6 μg/ml chloramphenicol and grown overnight at37° C. The following day 188 individual colonies were picked from eachSSL and used to inoculate 200 μl TB-Gly with 6 μg/ml chloramphenicol in96 well MTPs. The plates were grown overnight, and the mutationcontained in each clone was identified by sequencing. A culturerepresenting each of the 20 possible amino acid substitutions at eachposition in the enzyme was then transferred from the grown MTPs to a newMTP and all these collected variants were for screening.

Growth and Expression

Cultures inoculated from the MTPs of collected variants were grown indeep well plates with 600 μl Cal-18 (Ostergaard PR, Wilting R, LassenSF. 2010. Identification and characterization of a bacterial glutamicpeptidase. BMC Biochem. 11:47) supplemented with 6 μg/ml chloramphenicolat 37° C. for one day with shaking at 700 rpm. The wild type was alsoinoculated as reference in four wells on each microtiterplate. Aftergrowth, the plates were centrifuged to pellet the cells and the variantspresent in the supernatants were screened for improved stability.

EXAMPLES Example 1 Growth and Expression

The constructed variants were plated on LB agar supplemented with 6μg/ml chloramphenicol and grown for 37 ′C for one day. After growth,colonies were picked to individual wells of deep well 96-well microtiterplates or standard 96-well microtiter plates containing 600 μl or 200 ulCal18-2 (Ostergaard PR, Wilting R, Lassen SF. 2010. Identification andcharacterization of a bacterial glutamic peptidase. BMC Biochem. 11:47)broth, respectively, supplemented with 6 μg/ml chloramphenicol. The wildtype dispersin (SEQ ID NO: 1), was also inoculated as reference in fourwells on each microtiterplate. The deepwell microtiter plates were grownfor one day at 37 ′C with shaking at 700 rpm and the standard 96-wellmicrotiter plates grown for 3 days at 30° C. with shaking at 225 rpm.

Example 2a Testing the Variants for Stability Measurement ofThermostability at pH 8

The variants were made as a site-saturation library as described above.Each supernatant sample, grown as stated in example 1, was diluted20-fold in 50 mM Tris-HCI, pH 8.0. The sample was split in two parts:One part was incubated for 60 minutes at room temperature (21° C.,unstressed sample) whereas the second part was incubated in a PCRmachine for 60 minutes at 41° C. (stressed sample). After theincubation, 5 μl of the samples was transferred to the 384-well MTPcontaining 35 pl dispersin assay solution (45 mM citrate buffer pH 5added 0.5 mg/ml p-nitrophenyl-N-acetyl-β-D-glucosaminide). The 384-wellplate was then incubated at room temperature for 3 hours. Afterincubation, 40 μl stop solution (0.4 M Na₂CO₃) was added to the sampleand absorbance at 405 nm was read. The measurements were corrected forthe background by subtraction of the absorbance measurement obtained fora reference without dispersin. The residual activity (RA) for eachdispersin variant and the reference dispersin was calculated as theratio of background corrected activity between a stressed and anunstressed sample. These values were subsequently used to calculate theresidual activity ratio (RAR=RA_(varinant)/RA_(reference)), which wasused as selection criteria of hits with improved stability. Variantswith improved stability will have a RAR>1 as the RAR of the backbone inthis setup will by definition be 1.0. The result is shown in Table 1.

TABLE 1 Variant RAR Variant RAR Variant RAR Variant RAR D2V 1.23 Y124K1.22 S199L 1.20 A284T 3.32 S12A 2.58 D125R 3.36 A203G 3.06 A284L 3.01T17E 2.53 D125C 2.35 A203E 1.36 A284I 1.47 V18L 1.22 D125G 2.01 A203V1.10 S288K 4.43 K22M 2.17 D125H 1.58 N204L 2.02 V290I 1.43 S23I 1.65D125K 1.29 N204Y 1.72 K291R 1.36 S23C 1.45 D125Q 1.18 N204V 1.21 T296C1.18 S23A 1.45 Q135M 1.16 L205I 1.78 T303Q 4.45 S23T 1.41 D138R 2.44D207K 3.47 D304M 2.00 S23L 1.33 D138K 1.57 D207S 3.01 D304C 1.12 S23V1.24 D138N 1.32 D207C 2.06 L305M 4.54 S23E 1.18 D138Q 1.14 D207G 1.90L305N 1.19 L24I 2.68 S163P 1.67 S210T 1.32 S306C 1.48 V25R 1.31 L170D2.47 Q215K 3.95 K308D 6.09 Y30A 3.11 L170K 1.78 Q215R 2.59 K308E 5.11Y30L 1.85 L170S 1.43 S221N 1.22 K308A 3.83 A49W 2.38 L170H 1.33 N227T1.99 K308V 3.79 A49Y 1.89 D171E 3.61 N227K 1.50 K308Q 3.73 S56T 1.18D171K 3.26 Y244M 1.47 K308S 3.21 N59D 1.78 D171Y 2.33 Y244C 1.21 K308Y1.96 N59C 1.59 D171M 2.01 N252C 1.15 K308G 1.54 N59E 1.51 D171Q 1.57Q256E 1.54 K308L 1.50 N59R 1.29 D171L 1.37 Q256D 1.31 K308T 1.37 N59F1.25 I173C 1.41 Q262H 2.32 K308I 1.35 N59W 1.22 D174W 2.05 M263Q 1.13K309E 5.27 N59V 1.20 D174H 1.69 D264E 1.35 K309G 4.86 N62C 1.69 D174M1.33 Y265F 3.45 K309C 4.84 N62D 1.42 D174N 1.28 N267T 2.40 K309L 4.16T63C 1.38 F175Y 1.39 N267S 2.07 K309D 3.99 N68Q 1.11 Q178K 1.91 Y270F1.22 K309Q 3.97 S72D 2.10 I179T 4.04 H272M 3.49 K309N 3.71 S72E 1.69S181F 3.68 H272P 2.28 K309T 3.60 I74L 1.11 S181Q 3.41 H272I 1.87 K309A3.50 S77A 1.91 S181G 2.75 H272V 1.57 K309S 3.48 I82V 1.83 S181N 2.32N273W 1.13 K309M 2.25 L90F 3.44 S181C 1.68 K274H 2.27 K309H 2.23 E99Q1.76 S181K 1.59 F276A 4.23 K312E 3.79 L100S 2.05 E185A 4.31 F276N 3.10K312A 3.11 V106Y 1.53 E185R 1.50 F276K 2.24 K312Q 2.65 T114S 3.12 E185M1.44 F276C 2.03 K312S 2.39 T114C 1.31 E185V 1.28 I278V 1.59 K312W 2.05Y123W 1.92 S186K 2.88 S279N 2.85 K312L 1.33 Y124I 2.48 S186M 2.10 S279D1.21 K312N 1.22 Y124M 2.44 S186R 1.89 S279G 1.17 E314L 1.23 Y124H 2.14S186H 1.84 D280E 3.19 E314V 1.19 Y124R 2.09 K187G 1.21 D280C 1.17 E314I1.14 Y124V 1.98 Y188P 2.71 Y281P 4.70 L315I 2.42 Y124Q 1.97 E189V 1.12Y282N 1.65 R319A 1.21 Y124T 1.38 S199C 1.42 H283I 3.92 Y321F 1.58 N323R1.20

Table 1 shows that all the dispersin variants have improved RAR i.e. RARabove 1, and thus are more stable under the tested conditions comparedto the dispersin without the above substitutions e.g. compared to thedispersin shown in SEQ ID NO: 1.

Example 2b Testing the Variants for Stability Growth and Expression

The constructed variants and the backbone used for the variants weregrown in standard 96-well microtiter plates (200 ul broth/well). Theused broth for growth and expression was Cal18-2 (Ostergaard P R,Wilting R, Lassen S F. 2010. Identification and characterization of abacterial glutamic peptidase. BMC Biochem. 11:47) supplemented with 6ug/ml chloramphenicol. The microtiter plates were grown for 3 days at30° C. with shaking at 225 rpm. After growth, the plates werecentrifuged, and the supernatants were stressed and assayed forstability. All steps regarding growth, stress and assaying are done in96- or 384-well format microtiter plates.

Measurement of Stability in the Presence of 80% (v/v) Model A +/−1.0%(v/v) Protease

The supernatants were diluted 5-fold in concentrated Model A detergentcontaining 1.25% (v/v) protease (SEQ ID NO:6). After mixing, the sampleswere split in two parts. One part was incubated for 60 minutes at roomtemperature (21° C., unstressed sample) whereas the second part wasincubated in a PCR machine for 60 minutes at 56° C. (stressed sample).After incubation, the samples were diluted 8-fold in dilution buffer(100 mM Tris-HCI, 0.01% (v/v) TritonX-100, pH 8.0) before the activityof the unstressed and stressed samples was determined by transferring 5ul sample to a 384-well microtiter plate containing 35 ul assay solution(45 mM citrate buffer pH 5.0 supplemented with 1.0 mg/mlp-nitrophenyl-N-acetyl-β-D-glucosaminide (Sigma Aldrich). Afterincubation at room temperature for 12 hours, the reactions were stoppedby adding 40 μl stop solution (0.4 M Na₂CO₃) and the absorbances at 405nm (A405) read. For each sample the residual activity (RA) wascalculated as: RA=A405(stressed sample)/A405(unstressed sample). AllA405 measurements were corrected for the signal of a blank sample (nodispersin present) before calculating the RA. The RA was used tocalculate the half-life of the variants and the backbone: Half-life(minutes)=60(minutes) × Ln(0.5)/Ln(RA) of each variant and the backbone.For each variant the half-life improvement factor (HIF) are calculatedas: HIF=Half-life(variant)/Half-life(backbone). Variants with improvedstability will have a HIF>1 as the HIF of the backbone in this setupwill by definition be 1.0.

TABLE 2 Variant HIF Variant HIF Variant HIF Variant HIF D2L 1.49 V106M1.16 G164D 1.39 D230T 1.20 D2A 1.23 V106D 1.15 G164H 1.26 N233E 1.56 D2W1.21 V106K 1.15 G164S 1.25 N233D 1.54 D2V 1.20 V106A 1.14 G164E 1.24N233H 1.28 D2R 1.11 V106Q 1.13 G164V 1.11 N233Q 1.25 D2N 1.11 K107M 1.32V167Q 2.66 N233R 1.18 Q3L 3.59 K107W 1.30 V167A 1.38 N233W 1.11 Q3V 2.75K107L 1.23 V167W 1.36 W234R 1.36 Q3Y 2.32 K107C 1.21 V167P 1.32 G235A1.43 Q3M 2.13 K107V 1.19 V167D 1.26 G235S 1.42 Q3I 1.64 K107A 1.13 V167L1.22 G235V 1.40 Q3P 1.61 K107T 1.12 V167R 1.20 G235L 1.38 H15Y 2.20N110R 1.29 V167E 1.18 G235M 1.33 H15F 1.27 N110V 1.12 L170N 1.56 G235E1.25 T17V 1.55 N110M 1.11 L170R 1.55 G235F 1.24 T17R 1.31 D111V 1.57L170D 1.43 G235P 1.14 T17M 1.30 D111A 1.29 L170E 1.37 G235I 1.12 T17F1.30 D111R 1.21 L170K 1.26 G235W 3.10 T17E 1.28 D111N 1.19 L170F 1.26S237W 2.68 T17C 1.20 D111W 1.17 L170M 1.23 S237Y 1.66 T17W 3.50 D111M1.12 L170A 1.23 S237G 1.38 E19P 1.25 D111Q 1.11 L170W 1.20 S237M 1.37E19N 1.21 D111E 1.11 L170Y 1.17 S237N 1.33 E19K 1.10 V113T 1.13 L170V1.15 S237C 1.25 E19D 1.10 Y116R 1.18 L170Q 1.15 Y244E 1.59 K22A 1.31Y116D 1.14 L170P 1.12 L249Q 2.38 K22V 1.22 Y116N 1.14 D171L 1.35 L249R1.65 S23V 1.53 S117N 1.27 D171V 1.31 L249W 1.60 S23R 1.40 S117H 1.21D171A 1.18 L249H 1.41 S23T 1.34 S117D 1.18 D171W 1.13 L249K 1.14 S23A1.22 S117P 1.14 D171R 1.11 L249Y 1.11 D26M 1.13 S117* 1.12 D171C 1.10S251A 1.41 Y30* 1.60 E118G 1.32 D174V 1.43 S251L 1.30 Y30R 1.43 E118*1.18 N177M 1.12 S251R 1.22 Y30V 1.39 E118A 1.17 Q178R 1.26 S251W 1.14Y30D 1.31 E118L 1.14 Q178* 1.23 S251N 1.12 Y30A 1.24 E118D 1.12 Q178W1.23 G253D 1.48 Y30T 1.20 E119W 1.71 Q178K 1.16 G253W 1.16 Y30N 1.16E119G 1.22 Q178A 1.15 F254I 2.61 Y30L 1.15 T120W 1.76 S181W 1.24 F254L2.27 Y30M 1.14 T120M 1.55 S181C 1.23 F254M 1.60 G32R 1.70 T120V 1.24S181V 1.21 F254Y 1.52 G32M 1.32 T120L 1.16 S181D 1.18 Q256R 1.15 G32L1.23 T120I 1.14 S181T 1.15 N260Q 2.39 N34D 1.45 D122* 1.26 S181P 1.12N260L 1.80 N43* 1.24 D122R 1.22 G235N 1.32 N260C 1.68 N43L 1.20 H168N1.22 G235H 1.31 N260R 1.60 N43H 1.13 D122H 1.13 E185W 1.40 N260M 1.55E44* 1.24 Y124L 1.33 S186R 1.81 N260K 1.51 N45D 1.19 Y124N 1.21 S186D1.29 N260A 1.47 N45L 1.19 Y124W 1.19 S186E 1.23 N260V 1.41 N45V 1.10Y124C 1.18 S186N 1.22 N260* 1.37 Y52M 1.13 Y124R 1.16 S186Q 1.18 N260I1.26 Y52* 1.12 D125H 1.42 S186L 1.15 N260T 1.23 G54N 1.17 N126V 1.11S186K 1.13 N260E 1.23 G54L 1.13 R127L 1.26 S186W 1.12 N260W 1.18 G54M1.12 R127W 1.25 S186V 1.12 N260Y 1.14 S56W 1.11 R127Q 1.22 S186I 1.10E261A 1.34 S57W 1.21 R127M 1.20 K187R 1.31 E261D 1.21 E58N 1.30 R127D1.15 K187W 1.25 E261R 1.15 N59R 1.43 R127H 1.15 K187V 1.24 E261W 1.12N59A 1.43 R127K 1.14 K187C 1.22 E261* 1.11 N59C 1.39 V128L 1.22 K187D1.20 Q262F 2.05 N59M 1.38 V128D 1.15 K187S 1.14 Q262W 1.28 N59D 1.30V128A 1.14 K187G 1.12 Q262Y 1.26 N59W 1.14 V128C 1.14 E189L 1.23 Q262*1.22 N59F 1.11 V128T 1.12 E189W 1.13 M263L 1.12 T60V 1.19 D131V 1.15S199Y 1.27 M263K 1.11 N62D 1.38 Q135M 1.37 S199M 1.19 D264* 1.27 N62W1.17 Q135* 1.29 S199C 1.16 D264C 1.17 N62Q 1.14 Q135Y 1.16 S199L 1.14D264N 1.16 N62H 1.10 Q135A 1.14 E200W 1.53 W268E 1.79 T63V 1.34 Q135D1.13 E200D 1.20 W268R 1.26 T63L 1.32 Q135E 1.13 E200K 1.18 W268C 1.26T63N 1.31 Q135K 1.11 E200R 1.13 W268M 1.10 T63R 1.23 D138M 1.30 E200M1.12 A271G 1.27 T63D 1.13 D138V 1.28 E200F 1.11 A271D 1.16 K67L 1.17D138R 1.27 E200N 1.10 H272M 3.24 K67A 1.15 D138L 1.27 E200L 1.10 H272V1.85 N68L 1.14 D138W 1.25 A203V 1.37 H272D 1.84 L71R 1.25 D138S 1.14A203R 1.33 K274R 1.24 L71H 1.22 E139W 1.18 A203G 1.31 K274A 1.14 L71N1.20 D142W 1.48 A203W 1.26 I278V 2.28 L71W 1.20 D142R 1.24 A203P 1.25I278Q 1.41 L71V 1.11 Y145V 1.51 A203C 1.21 I278A 1.40 S72T 1.39 Y145N1.34 A203L 1.20 I278N 1.37 S72G 1.34 Y145* 1.22 A203S 1.17 I278K 1.17S72C 1.30 Y145H 1.14 A203M 1.17 S279D 3.41 S72M 1.30 Y145L 1.11 A203Q1.14 S279C 1.52 S72F 1.24 P147L 1.36 A203T 1.13 S279N 1.51 S72N 1.24P147D 1.27 A203D 1.12 S279G 1.46 S72Y 1.23 P147A 1.26 N204V 1.22 S279E1.19 S72* 1.23 P147V 1.25 N204W 1.19 Y281P 4.46 S72R 1.17 P147G 1.21N204M 1.15 Y281H 2.40 S72E 1.16 P147R 1.17 D207N 1.72 Y281A 1.45 S72I1.14 P147S 1.17 D207V 1.54 Y281K 1.34 D79V 1.11 P147M 1.15 D207R 1.33Y281N 1.28 K80N 1.36 P147T 1.14 D207Q 1.27 Y281C 1.16 K80R 1.31 P147C1.14 D207E 1.15 Y281* 1.13 K80W 1.23 P147F 1.11 D207W 1.14 Y282E 1.10K80E 1.23 K148L 1.20 D207K 1.13 A284V 1.95 K80* 1.21 K148D 1.19 D207A1.12 A284N 1.29 K80L 1.14 K148A 1.18 S208A 1.59 A284P 1.16 K80V 1.11K148V 1.16 S208V 1.35 A284T 1.12 K80Q 1.10 F149N 1.28 S208C 1.26 T287N1.15 K80H 1.10 F149M 1.25 S208W 1.23 S288D 1.56 D81G 1.44 F149L 1.18S208D 1.20 S288N 1.49 D81A 1.36 E150N 1.48 S208T 1.18 K291L 1.13 D81L1.33 E150R 1.35 S208Q 1.13 K291R 1.12 D81W 1.29 E150D 1.31 S208G 1.12K291V 1.11 D81T 1.27 E150K 1.27 S208L 1.12 E300D 1.45 D81V 1.26 E150M1.21 Q215L 1.24 E300A 1.17 D81R 1.19 E150H 1.20 Q215R 1.17 H301N 1.95D81S 1.19 E150W 1.18 Q215M 1.16 H301C 1.66 D81N 1.14 E150V 1.18 Q215*1.11 H301R 1.46 E99R 1.12 E150Y 1.17 S217V 1.84 T303A 1.71 K103R 1.45E150L 1.15 T218Q 1.83 T303C 1.29 K103V 1.35 E150A 1.12 T218R 1.27 T303K1.24 K103A 1.20 G151A 1.45 T218A 1.25 T303G 1.24 K104N 1.29 G151P 1.45T218V 1.20 T303Q 1.22 K104W 1.22 G151W 1.27 T218L 1.10 T303W 1.21 D105N1.11 G151D 1.25 S221N 1.17 T303R 1.18 V106L 1.36 G151L 1.22 G222D 1.16K312M 1.26 V106Y 1.28 G151S 1.21 E224A 1.44 K312W 1.10 V106H 1.26 G151N1.16 E224P 1.28 L315V 1.85 V106E 1.25 G151C 1.15 D230N 1.71 L315I 1.36V106W 1.24 K152D 1.15 D230R 1.53 V106N 1.23 K152L 1.12 D230* 1.42 V106R1.20 K152R 1.10 D230W 1.28

Table 2 shows that all the dispersin variants have improved HIF i.e. HIFabove 1, and thus are more stable under the tested conditions comparedto the dispersin without the above alterations e.g. compared to thedispersin shown in SEQ ID NO: 1.

Example 3 Generation of SSLs and Screening Resulting Libraries as PoolsConstruction of SSL

A site-saturation library was constructed for each amino acid positionin the enzyme and the expected diversity was confirmed by sequencing ofthe pools of transformants. The pools of grown transformants were storedat −80° C. until use.

Growth and Expression

Cultures inoculated from the pools of transformants were grown in deepwell plates with 600 μl Cal-18 (Ostergaard P R, Wilting R, Lassen S F.2010. Identification and characterization of a bacterial glutamicpeptidase. BMC Biochem. 11:47) supplemented with 6 μg/ml chloramphenicolat 37° C. for one day with shaking at 700 rpm. The wild type was alsoinoculated as reference in four wells on each microtiterplate. Aftergrowth, the plates were centrifuged to pellet the cells and the variantspresent in the supernatants were screened for improved stability.

Testing Dispersin Variants for Stability

Each supernatant sample was diluted 20-fold in 50 mM Tris-HCI, pH 8.0.The sample was split in two: 5 μl is transferred to a 384-well MTP(unstressed reference) and 10 μl is transferred to 384-well PCR plate.The PCR plate was stressed in a PCR machine at 47° C. for 30 min, whilethe unstressed reference plate was kept at room temperature (21° C.).After the incubation, 5 μl of the stressed sample was transferred to the384-well MTP before assaying the activity in all samples by usage ofp-nitrophenyl-N-acetyl-β-D-glucosaminide (pNP-NAG).

Dispersin Activity Assay

For the dispersin activity measurement, the prepared 384-well microtiterplate containing 5 μl samples was added 35 μl dispersin assay solution(45 mM citrate buffer pH 5 added 0.5 mg/mlp-nitrophenyl-N-acetyl-β-D-glucosaminide). The 384-well plate was thenincubated at room temperature for 3 hours. After incubation, 40 μl stopsolution (0.4 M Na₂CO₃) was added to the sample and absorbance at 405 nmwas measured. The obtained activity values were corrected for thebackground by subtraction of the absorbance measurement obtained for areference without dispersin. The residual activity (RA) for each pool ofdispersin variants and the reference dispersin was calculated as theratio of background corrected activity between a stressed and anunstressed sample. These values were subsequently used to calculate theresidual activity ratio (RAR_(pool)=RA_(pool)/RA_(reference)), which wasused to prioritize of libraries in terms of likelihood of containingstabilized variant(s).

SSL Hit Selection

The result obtained for the pool will be the average of all variantscontained in the pool and the likelihood of finding improved candidatestherefore increase significantly the higher the value of the ratio(RAR_(pool)). SSLs with highest ratio and above that determined for thereference (i.e. RAR_(pool)>1) are deemed to be most interesting. In thisparticular example, six SSL pools with RAR_(pool)>2 were selected toscreen for improved properties amongst isolated clones representedtherein.

Plating, Regrow, Rescreen and Sequence

The selected variant pools (hits) were plated on LB agar supplementedwith 6 μg/ml chloramphenicol and grown for 37° C. for one day. Aftergrowth, colonies were picked to individual wells of 96-well deep wellplates containing 600 μl Cal18-2 (Ostergaard PR, Wilting R, Lassen SF.2010. Identification and characterization of a bacterial glutamicpeptidase. BMC Biochem. 11:47) broth supplemented with 6 μg/mlchloramphenicol. Dispersin (SEQ ID NO: 1), was also inoculated asreference in four wells on each microtiterplate. The microtiter plateswere grown for one day at 37° C. with shaking at 700 rpm. After growth,the supernatants were screened for residual activity after stressingthem for 30 minutes at 47° C. in 50 mM Tris-HCI, pH 8.0. The followingcriteria was used for qualification of the collected data. Thebackground corrected activity obtained for the unstressed referenceshould be at least 0.25. The criteria for selection of hits was basedupon the RA obtained for WT, which was determined to 0.1. A sample wasconsidered a hit if its RA_(varinant)/RA_(reference) equal to RAR>1.

TABLE 3a Mutations of position RA_(pool): 30 min. 47° C. in SEQ ID NO: 150 mM Tris-HCl. pH 8.0 RAR_(pool) SEQ ID NO: 1 0.12 1.00 2 0.13 1.11 30.24 1.98 20 0.21 1.74 23 0.13 1.09 24 0.16 1.37 29 0.15 1.26 30 0.151.25 31 0.22 1.82 33 0.25 2.11 36 0.15 1.26 42 0.19 1.58 43 0.20 1.64 450.22 1.82 59 0.27 2.27 72 0.12 1.03 73 0.17 1.41 114 0.12 0.98 124 0.312.55 125 0.13 1.09 134 0.14 1.13 135 0.13 1.04 138 0.21 1.77 171 0.151.27 174 0.17 1.39 181 0.26 2.18 185 0.16 1.31 186 0.18 1.49 199 0.141.17 207 0.20 1.66 215 0.15 1.26 216 0.17 1.39 225 0.23 1.89 227 0.252.10 230 0.18 1.52 243 0.15 1.22 244 0.22 1.80 249 0.16 1.30 252 0.151.21 253 0.21 1.79 254 0.12 0.98 256 0.12 0.97 261 0.12 1.03 262 0.120.96 265 0.16 1.36 272 0.22 1.85 276 0.31 2.56 294 0.12 0.99 303 0.121.01 308 0.37 3.05 309 0.55 4.59 312 0.40 3.36 322 0.15 1.25

Table 3a shows the subset of SSL libraries having improved RAR_(pool)i.e. RAR_(pool) above 1, and thus contain a mixture of substitutionsgiving more stability on average under the tested conditions compared tothe dispersin without the substitutions e.g. compared to the dispersinshown in SEQ ID NO: 1

The experiment above was repeated by screening pools made using astabilized enzyme variant (SEQ ID NO: 17) as template during SSLconstruction but the residual activity ratio (RAR) was measured in modeldetergent A, with the presence of a protease (SEQ ID NO: 16). Theresults are shown in table 3a* below.

TABLE 3a* Mutations compared to RAR_(pool): 60 min. 64° C. 10 g/L modelSEQ ID NO: 17 A detergent added 1 v/v % protease SEQ ID NO: 17 1.00 11.54 3 8.53 6 1.33 57 1.21 73 1.76 75 1.30 90 1.86 94 2.14 116 1.40 1411.11 142 1.10 144 1.07 147 1.84 149 1.34 169 1.35 170 1.14 193 6.30 2051.94 217 1.71 225 5.32 232 6.95 233 1.17 241 1.96 243 1.73 253 1.39 2543.15 261 1.29 267 2.61 272 33.88 276 1.53 277 6.31 278 1.17 279 1.98 2814.19 284 1.34 289 3.89 296 1.30 297 1.67 299 1.60 301 1.09

The experiment above was repeated by screening pools made using (SEQ IDNO: 1) as template during SSL construction and the residual activityratio (RAR) was measured in model detergent A. The results are shown intable 3b below.

TABLE 3b Mutations compared to RA: 30 min, 47° C., 50 mM SEQ ID NO: 1Tris-HCl, pH 8.0 RAR SEQ ID NO: 1 0.13 1.00 Y124A 0.22 1.70 Y124C 0.634.82 Y124F 0.44 3.38 Y124K 0.41 3.13 Y124L 0.44 3.39 Y124M 0.66 5.06Y124R 0.66 5.03 Y124S 0.16 1.25 Y124V 0.59 4.52 Y124W 0.37 2.82 N227A0.80 6.09 N227K 0.92 7.04 N227Q 0.93 7.11 N227R 1.11 8.49 N227S 0.796.02 N227T 1.23 9.43 F276A 0.90 6.86 F276C 0.93 7.14 F276G 0.24 1.85F276K 0.44 3.35 F276L 0.17 1.31 F276M 0.39 2.97 F276P 0.38 2.94 F276S0.35 2.67 F276V 0.23 1.74 F276W 0.38 2.88

Table 3b shows dispersin variants having improved RAR i.e. RAR above 1,and thus are more stable under the tested conditions compared to thedispersin without the above substitutions e.g. compared to the dispersinshown in SEQ ID NO: 1

The experiment above was repeated by screening selected positions fromSSLs made using a stabilized enzyme variant (SEQ ID NO: 17) as templatebut the residual activity ratio (RAR) was measured in model detergent A,with the presence of a protease (SEQ ID NO: 16). The results are shownin table 3b* below.

TABLE 3b* Mutations compared to RAR: 60 min. 63° C. 10 g/L model A SEQID NO: 17 detergent added 1 v/v % protease SEQ ID NO: 17 1 Q3F 9.72 Q3I8.24 Q3L 8.80 Q3T 8.03 Q3V 5.89 S225G 6.79 E232D 9.03 E232V 9.90 H272M16.45 H272P 11.91 H272V 12.48 H272W 9.03 S279G 3.60 Y281K 7.64 Y281P11.25 Y281R 5.80

The experiment above was repeated by screening pools made using SEQ IDNO: 1 as template during SSL construction and the residual activityratio (RAR) was measured in model detergent A. The results are shown intable 3c below.

TABLE 3c Mutations compared to RA: 30 min, 47° C., 50 mM SEQ ID NO: 1Tris-HCl, pH 8.0 RAR SEQ ID NO: 1 0.17 1.00 K308A 0.38 2.23 K308D 0.543.19 K308E 0.62 3.64 K308G 0.26 1.53 K308S 0.34 2.02 K308T 0.28 1.63K308V 0.17 1.02 K308Y 0.17 1.01 K309A 0.35 2.07 K309C 0.53 3.10 K309E0.56 3.30 K309G 0.49 2.88 K309H 0.23 1.38 K309I 0.18 1.07 K309L 0.191.11 K309N 0.35 2.08 K309Q 0.47 2.78 K309S 0.35 2.06 K309T 0.34 1.98K312A 0.43 2.52 K312E 0.66 3.93 K312L 0.27 1.59 K312N 0.31 1.85 K312Q0.48 2.85 K312S 0.45 2.65

Table 3c shows dispersin variants having improved RAR i.e. RAR above 1,and thus are more stable under the tested conditions compared to thedispersin without the above substitutions e.g. compared to the dispersinshown in SEQ ID NO: 1

Example 4 Combination Variants with 1-3 Mutations Growth and Expression

The constructed variants and the backbone used for the variants weregrown in standard 96-well microtiter plates 200 ul Cal18-2 broth(Ostergaard P R, Wilting R, Lassen S F. 2010. Identification andcharacterization of a bacterial glutamic peptidase. BMC Biochem. 11:47)supplemented with 6 ug/ml chloroampenicol broth/well. The microtiterplates were grown for 3 days at 30° C. with shaking at 225 rpm. Aftergrowth, the plates were centrifuged, and the supernatants were stressedand assayed for stability. All steps regarding growth, stress andassaying are done in 96- or 384-well format microtiter plates.

Measuring Stability at Elevated Temperature

Each supernatant sample was diluted 10-fold in stress buffer (100 mMTris-HCI, 0.01% (v/v) TritonX-100, pH 8.0. After mixing, the sampleswere split in four and two samples incubated at room temperature (21°C., unstressed samples) and the other two samples (stressed samples)incubated in a PCR machine for 120 minutes at 41° C. and 46° C.,respectively. After incubation, the activity of the un-stressed andstressed samples was determined by transferring 5 ul of each sample to a384-well microtiter plate containing 35 ul assay solution (45 mM citratebuffer pH 5.0 supplemented with 1.0 mg/mlp-nitrophenyl-N-acetyl-β-D-glucosaminide (Sigma Aldrich). Afterincubation at room temperature for 3 hours, the reactions were stoppedby adding 40 μl stop solution (0.4 M Na₂CO₃) and the absorbances at 405nm (A405 nm) read. For each sample the residual activity (RA) wascalculated as: RA=A405(stressed sample)/A405(unstressed sample). AllA405 nm measurements were corrected for the signal of a blank sample (nodispersin present) before calculating the RA. A variant is improved instability if RA(variant)>RA(backbone).

TABLE 4 RA (residual activity) Number of 41° C., 46° C., substitutionsMutations 120 min 120 min 0 SEQ ID NO: 1 0.07 0.01 1 F276A 0.90 0.17 1K308E 0.92 0.10 1 K309E 0.94 1 K312E 0.91 0.13 1 N227T 0.78 0.05 1 N252P0.45 0.02 1 N267T 0.56 0.01 1 Q215K 0.63 0.03 1 S163P 0.48 0.02 2F276A + K308E 0.98 0.98 2 F276A + K309E 1.00 1.00 2 K308E + K309E 0.970.97 2 K308E + K312E 0.99 0.88 2 K309E + K312E 1.00 0.93 2 N227T + F276A1.00 0.98 2 N227T + K308E 0.97 0.94 2 N227T + K312E 0.99 0.94 2 N227T +N252P 1.00 0.15 2 N227T + N267T 0.94 0.40 2 N252P + F276A 0.98 0.97 2N252P + K308E 1.00 0.26 2 N252P + K309E 0.99 0.81 2 N252P + N267T 0.750.02 2 N267T + F276A 0.97 0.63 2 N267T + K308E 1.00 0.32 2 N267T + K309E0.98 0.79 2 N267T + K312E 0.97 0.59 2 Q215K + F276A 1.00 0.79 2 Q215K +K308E 0.94 0.62 2 Q215K + K309E 0.98 0.75 2 Q215K + K312E 0.97 0.71 2Q215K + N227T 0.98 0.60 2 Q215K + N252P 0.83 0.03 2 Q215K + N267T 0.960.05 2 S163P + F276A 0.97 0.79 2 S163P + K308E 0.96 0.75 2 S163P + K309E0.96 0.83 2 S163P + K312E 0.97 0.88 2 S163P + N252P 0.96 0.05 2 S163P +N267T 0.64 0.03 2 S163P + Q215K 0.91 0.14 3 F276A + K308E + K309E 0.970.96 3 F276A + K308E + K312E 0.99 1.00 3 F276A + K309E + K312E 1.00 1.003 K308E + K309E + K312E 1.00 0.99 3 N227T + F276A + K309E 0.99 0.97 3N227T + F276A + K312E 0.99 0.98 3 N227T + K308E + K309E 0.99 1.00 3N227T + K308E + K312E 1.00 1.00 3 N227T + K309E + K312E 1.00 0.99 3N227T + N252P + K308E 1.00 1.00 3 N227T + N252P + K309E 0.99 0.99 3N227T + N252P + K312E 1.00 1.00 3 N227T + N252P + N267T 1.00 1.00 3N227T + N267T + F276A 1.00 0.99 3 N227T + N267T + K308E 0.99 0.97 3N227T + N267T + K309E 0.97 0.96 3 N227T + N267T + K312E 1.00 0.89 3N252P + F276A + K308E 0.96 0.97 3 N252P + F276A + K309E 1.00 0.99 3N252P + F276A + K312E 0.98 0.97 3 N252P + K308E + K309E 0.99 0.93 3N252P + K308E + K312E 0.97 0.95 3 N252P + N267T + F276A 1.00 0.97 3N252P + N267T + K308E 1.00 1.00 3 N252P + N267T + K309E 1.00 1.00 3N252P + N267T + K312E 0.99 0.98 3 N267T + F276A + K308E 1.00 1.00 3N267T + F276A + K309E 0.99 1.00 3 N267T + F276A + K312E 0.98 0.97 3N267T + K308E + K309E 1.00 1.00 3 N267T + K308E + K312E 0.97 0.98 3N267T + K309E + K312E 1.00 0.98 3 Q215K + F276A + K308E 0.98 0.92 3Q215K + F276A + K312E 1.00 1.00 3 Q215K + K308E + K312E 0.99 0.94 3Q215K + K309E + K312E 0.98 0.98 3 Q215K + N227T + F276A 1.00 1.00 3Q215K + N227T + K308E 0.99 0.99 3 Q215K + N227T + K309E 0.97 0.97 3Q215K + N227T + K312E 0.96 0.96 3 Q215K + N227T + N252P 0.96 0.78 3Q215K + N227T + N267T 0.96 0.86 3 Q215K + N252P + F276A 1.00 0.96 3Q215K + N252P + K309E 0.98 0.93 3 Q215K + N252P + K312E 0.99 0.95 3Q215K + N252P + N267T 0.97 0.23 3 Q215K + N267T + F276A 0.99 0.97 3Q215K + N267T + K312E 1.00 0.96 3 S163P + F276A + K308E 1.00 1.00 3S163P + F276A + K312E 0.98 0.96 3 S163P + K308E + K309E 0.98 0.97 3S163P + K308E + K312E 1.00 1.00 3 S163P + K309E + K312E 0.99 1.00 3S163P + N227T + F276A 0.96 0.96 3 S163P + N227T + K309E 1.00 1.00 3S163P + N227T + N252P 1.00 0.98 3 S163P + N227T + N267T 0.97 0.86 3S163P + N252P + F276A 1.00 0.99 3 S163P + N252P + K308E 0.97 0.91 3S163P + N252P + K309E 0.99 0.98 3 S163P + N252P + K312E 0.98 0.96 3S163P + N252P + N267T 0.97 0.57 3 S163P + N267T + K308E 0.91 0.90 3S163P + N267T + K309E 0.99 0.99 3 S163P + N267T + K312E 0.98 0.96 3S163P + Q215K + F276A 0.99 0.95 3 S163P + Q215K + K308E 1.00 0.94 3S163P + Q215K + K309E 0.63 0.95 3 S163P + Q215K + K312E 0.94 0.95 3S163P + Q215K + N227T 0.95 0.90 3 S163P + Q215K + N252P 0.94 0.38

Table 4 shows the effect of adding several substitutions compared to SEQID NO: 1. It is shown that the single stabilizing substitutions can becombined to generate a variant which is more stable or on par to thecorresponding variant with one less substitution.

Example 5 Combination Variants with 4-9 Mutations Growth and Expression

The constructed variants and the backbone used for the variants weregrown in either 96-deep-Well (600 ul broth/well) or standard 96-wellmicrotiter plates (200 ul broth/well). The used broth for growth andexpression was Cal18-2 (Ostergaard P R, Wilting R, Lassen S F. 2010.Identification and characterization of a bacterial glutamic peptidase.BMC Biochem. 11:47) supplemented with 6 ug/ml chloramphenicol. Theplates were grown for 1 day at 37° C. with shaking at 700 rpm (96-welldeepwell plates) or for 3 days at 30° C. with shaking at 225 rpm(standard 96-well microtiter plates). After growth, the plates werecentrifuged, and the supernatants were stressed and assayed forstability. All steps regarding growth, stress and assaying are done in96- or 384-well format microtiter plates.

Measurement of Stability in the Presence of 0.9 g/L Model A +/−0.9%(v/v) Protease

Each supernatant sample was diluted 10-fold in stress buffer (100 mMTris-HCI, 0.01% (v/v) TritonX-100, pH 8.0 supplemented with 1 g/L ModelA detergent). In the cases with applied protease stress, the stressbuffer was also added 1% (v/v) protease (SEQ ID NO:7). After mixing, thesample was split into an unstressed sample which was incubated at roomtemperature (21° C.) and a stressed sample which was incubated in a PCRmachine for 30 minutes at the selected stress temperatures (50° C., 51°C., 54° C., 57° C. and/or 60° C.). After the incubation, the activity ofthe un-stressed and stressed sample was determined by transferring 5 ulof each sample to a 384-well microtiter plate containing 35 ul assaysolution (45 mM citrate buffer pH 5.0 supplemented with 1.0 mg/mlp-nitrophenyl-N-acetyl-β-D-glucosaminide (Sigma Aldrich). After 3 hourincubation at room temperature, the reactions were stopped by adding 40μl stop solution (0.4 M Na₂CO₃) and the absorbances at 405 nm (A405)read. For each sample the residual activity (RA) was calculated as:RA=A405(stressed sample)/A405(unstressed sample). All A405 measurementswere corrected for the signal of a blank sample (no dispersin present)before calculating the RA. A variant stabilized at the tested stressconditions is defined as a variant having a RA higher than the usedbackbone. All variants were tested in 30 min in 0.9 g/L Model Adetergent.

TABLE 5 RA RA (residual (residual activity) Number of activity) 50° C. +0.9% substitutions Substitutions compared to SEQ ID NO: 1 50° C.Protease 0 SEQ ID NO: 1 <0.05 <0.05 4 N227T + F276A + K308E + K309E 0.780.37 4 N227T + F276A + K308E + K312E 0.87 0.44 4 N227T + F276A + K309E +K312E 0.89 0.42 4 N227T + N267T + F276A + K308E 0.76 0.38 4 N227T +N267T + F276A + K309E 0.86 0.45 4 N227T + N267T + F276A + K312E 0.820.45 4 S163P + N227T + F276A + K308E 0.55 0.39 5 N227T + N267T + F276A +K308E + 0.92 0.54 K309E 5 N227T + N267T + F276A + K308E + 0.94 0.51K312E 5 N227T + N267T + F276A + K309E + 0.92 0.50 K312E 5 S163P +N227T + F276A + K308E + 0.90 0.74 K312E 5 N227T + F276A + K308E +K309E + 0.92 0.53 K312E 6 N227T + N267T + F276A + K308E + 0.94 0.53K309E + K312E

TABLE 6 RA RA (residual (residual activity) activity) Number of 51° C.0.9% 54° C. 0.9% substitutions Substitutions compared to SEQ ID NO: 1Protease Protease 5 N227T + F276A + K308E + K309E + K312E 0.36 0.24 6N227T + N267T + F276A + K308E + K309E + K312E 0.35 0.18 6 N227T +N252P + F276A + K308E + K309E + K312E 0.49 0.25 6 Q215K + N227T +F276A + K308E + K309E + K312E 0.50 0.36 6 S163P + N227T + F276A +K308E + K309E + K312E 0.71 0.63 7 N227T + N252P + N267T + F276A +K308E + K309E + K312E 0.41 0.26 7 Q215K + N227T + N252P + F276A +K308E + K309E + K312E 0.45 0.31 7 Q215K + N227T + N267T + F276A +K308E + K309E + K312E 0.51 0.32 7 S163P + N227T + N252P + F276A +K308E + K309E + K312E 0.80 0.74 7 S163P + N227T + N267T + F276A +K308E + K309E + K312E 0.75 0.72 7 S163P + Q215K + N227T + F276A +K308E + K309E + K312E 0.76 0.72 8 Q215K + N227T + N252P + N267T +F276A + K308E + K309E + 0.41 0.31 K312E 8 S163P + N227T + N252P +N267T + F276A + K308E + K309E + 1.02 0.81 K312E 8 S163P + Q215K +N227T + N252P + F276A + K308E + K309E + 0.81 0.77 K312E 8 S163P +Q215K + N227T + N267T + F276A + K308E + K309E + 0.81 0.72 K312E 9S163P + Q215K + N227T + N252P + N267T + F276A + K308E + 0.81 0.78K309E + K312E

TABLE 7 RA RA (residual (residual activity) activity) Number of 57° C.0.9% 60° C. 0.9% substitutions Substitutions compared to SEQ ID NO: 1Protease Protease 5 N227T + F276A + K308E + K309E + K312E 0.09 6 N227T +N267T + F276A + K308E + K309E + K312E 0.09 6 N227T + N252P + F276A +K308E + K309E + K312E 0.13 0.11 6 Q215K + N227T + F276A + K308E +K309E + K312E 0.16 0.10 6 S163P + N227T + F276A + K308E + K309E + K312E0.30 0.11 7 N227T + N252P + N267T + F276A + K308E + K309E + 0.10 0.08K312E 7 Q215K + N227T + N252P + F276A + K308E + K309E + 0.20 0.17 K312E7 Q215K + N227T + N267T + F276A + K308E + K309E + 0.14 0.10 K312E 7S163P + N227T + N252P + F276A + K308E + K309E + 0.54 0.37 K312E 7S163P + N227T + N267T + F276A + K308E + K309E + 0.44 0.27 K312E 7S163P + Q215K + N227T + F276A + K308E + K309E + 0.50 0.30 K312E 8Q215K + N227T + N252P + N267T + F276A + K308E + 0.15 0.11 K309E + K312E8 S163P + N227T + N252P + N267T + F276A + K308E + 0.60 0.40 K309E +K312E 8 S163P + Q215K + N227T + N252P + F276A + K308E + 0.66 0.46K309E + K312E 8 S163P + Q215K + N227T + N267T + F276A + K308E + 0.570.45 K309E + K312E 9 S163P + Q215K + N227T + N252P + N267T + F276A +0.58 0.46 K308E + K309E + K312E

Table 5, 6 and 7 shows the effect of adding several substitutionscompared to SEQ ID NO: 1. It is shown that the single stabilizingsubstitutions can be combined to generate a variant which is more or onpar in stability as the corresponding variant with one lesssubstitution.

Example 6 Testing for Thermostability Growth and Expression

The variants were grown and expressed as described in the previousexamples and purified as described below.

Purification

The culture broth was centrifuged (26000×g, 20 min) and the supernatantwas carefully decanted from the precipitate. The supernatant wasfiltered through a Nalgene 0.2 μm filtration unit to remove the rest ofthe Bacillus host cells. The 0.2 μm filtrate was transferred to 20 mMMES/NaOH, pH 6.0 on a G25 sephadex column (from GE Healthcare). The G25transferred solution was applied to a SOURCE Q column (from GEHealthcare) equilibrated in 20 mM MES/NaOH, pH 6.0. After washing thecolumn extensively with the equilibration buffer, the protease waseluted with a linear NaCI gradient (0→1.0 M NaCI) in the same bufferover five column volumes. Fractions were collected during elution andthe collected fractions were analysed by SDS-PAGE. Fractions, where onlyone band was seen after coomassie staining, were pooled as the purifiedpreparation and was used for further experiments.

Measurement of Stability

Nano differential scanning fluorimetry (nDSF) was utilized to determinethermal stability. Dispersin variants for nDSF were diluted to ˜0.2mg/mL (>10-fold dilution) in 50 mM 2-(N-Morpholin)ethansulfonsyre (MES),50 mM glycine, 50 mM acetic acid, pH 7 or 50 mM MES, 50 mM glycine, 50mM acetic acid, pH 7 and x (x=2 or 50) g/L Model A (model detergent A).Sample dilution was made in 96-well microtiter plates and transferred to384-well microplates. The instruments utilized for nDSF experiments,were either a Prometheus NT.48 or Prometheus NT.Plex with autosamplerfrom Nanotemper. With the Prometheus NT.48, the samples were loadedmanually using single capillaries (PR-0002), while samples were loadedin the Prometheus NT.Plex using capillary chips (PR-AC002). Theexperiments were conducted from 20 to 95° C. with a temperature gradientof 3.3° C./min. The melting temperatures (Tm-values) were obtained frompeak values derived from the first-derivative of the signal trace(350/330 nm fluorescence ratio or 330 nm fluorescence) usingPR.ThermControl software.

TABLE 8 Thermostability for variants Tm-values (° C.) 0 g/L 2 g/L 50 g/LSubstitutions compared to SEQ ID NO: 1 Model A Model A Model A SEQ IDNO: 1 50.53 40.94 F276A 55.22 53.86 52.36 K308E 54.14 K309E 55.48 52.7546.62 K312E 54.96 52.23 N227T 53.11 41.30 N227T + F276A 58.00 55.9949.87 N227T + F276A + K308E 63.38 62.25 58.99 N227T + F276A + K309E64.28 64.05 60.35 N227T + F276A + K312E 63.71 63.78 60.70 N227T +F276A + K308E + K309E 68.45 67.73 65.24 N227T + F276A + K308E + K312E67.19 66.32 63.88 N227T + F276A + K309E + K312E 67.37 66.9 64.57 N227T +N267T + F276A + K308E 64.71 55.32 63.10 N227T + N267T + F276A + K309E66.45 66.79 64.72 N227T + N267T + F276A + K312E 65.86 66.59 65.07N227T + F276A + K308E + K309E + K312E 68.42 67.96 66.06 N227T + N267T +F276A + K308E + K309E 70.36 69.86 68.2 N227T + N267T + F276A + K308E +K312E 69.33 68.89 66.83 S163P + N227T + F276A + K308E + K312E 67.5966.78 63.81 N227T + N267T + F276A + K308E + K309E + K312E 70.53 70.5468.88 S163P + Q215K + N227T + N252P + N267T + F276A 76.12 75.87 73.98S163P + N227T + N252P + F276A + K308E + 72.74 72.69 70.67 K309E + K312ES163P + N227T + N267T + F276A + K308E + 71.27 71.04 68.87 K309E + K312ES163P + Q215K + N227T + F276A + K308E + 71.42 68.59 K309E + K312ES163P + Q215K + N227T + N252P + N267T + 70.63 69.74 66.42 F276A + K308ES163P + Q215K + N227T + N252P + N267T + 73.46 73.41 69.96 F276A + K309ES163P + Q215K + N227T + N252P + N267T + 72.49 71.59 69.04 F276A + K312EQ215K + N227T + N252P + N267T + F276A + 75.79 75.53 74.29 K308E +K309E + K312E S163P + N227T + N252P + N267T + F276A + 74.87 74.82 73.40K308E + K309E + K312E S163P + Q215K + N227T + N252P + F276A + 75.1075.02 73.01 K308E + K309E + K312E S163P + Q215K + N227T + N252P +N267T + 76.61 75.93 73.57 F276A + K308E + K309E S163P + Q215K + N227T +N252P + N267T + 75.74 75.5 73.56 F276A + K309E + K312E S163P + Q215K +N227T + N252P + N267T + 72.05 71.6 68.52 K308E + K309E + K312E S163P +Q215K + N227T + N267T + F276A + 73.07 72.92 70.98 K308E + K309E + K312ES163P + Q215K + N252P + N267T + F276A + 72.68 72.66 70.85 K308E +K309E + K312E S163P + Q215K + N227T + N252P + N267T + 76.77 76.72 75.07F276A + K308E + K309E + K312E

TABLE 8a Tm-values (° C.) 0 g/L 2 g/L 50 g/L Substitutions compared toSEQ ID NO: 1 Model A Model A Model A SEQ ID NO: 1 50.53 40.94 N.D.S186R + N227T + E232D + K308E 59.60 57.39 50.18 S163P + Q215K + N227T +N252P + N267T + F276A + K308Q + K309E + K312E 65.25 65.08 59.92 S163P +Q215K + N227T + N252P + N267T + F276A + K308E + K309E + K312Q 76.8076.42 74.66 Q3F + A49W + N59E + S163P + S186R + N227T + E232D + N252P +F276A + 77.70 77.49 76.00 S279G + K308E + K309E + K312E A49W + N59E +S163P + S186R + S225G + N227T + E232D + N252P + F276A + 77.84 77.9376.16 S279G + K308E + K309E + K312E Q3F + T17W + A49W + N59E + S163P +S186R + S225G + N227T + G235W + 78.25 77.67 74.95 N252P + H272P +Y281P + K308Q + K309E + K312Q A49W + N59E + S163P + S186R + N227T +E232D + N252P + H272V + F276A + 78.39 78.31 76.49 S279G + K308E +K309E + K312E Q3F + A49W + N59E + S163P + S186R + S225G + N227T +N252P + F276A + 78.65 78.33 76.84 S279G + K308E + K309E + K312E A49W +N59E + S163P + S186R + N227T + E232D + N252P + F276A + S279G + 78.6878.13 76.54 Y281P + K308E + K309E + K312E A49W + N59E + S163P + S186R +S225G + N227T + N252P + F276A + S279G + 79.16 79.12 77.89 Y281P +K308E + K309E + K312E Q3F + A49W + N59E + S163P + S186R + S225G +N227T + 79.71 79.56 78.05 E232D + N252P + F276A + S279G + K308E +K309E + K312E Q3F + S163P + S186R + Q215K + N227T + E232D + N252P +80.17 79.52 78.43 N267T + F276A + S279G + K308E + K309E + K312E H15Y +A49W + N59E + S163P + S186R + S225G + N227T + E232D + G235W + 80.1980.20 78.91 N252P + N260Q + H272P + S279D + Y281P + K308Q + K309E +K312Q Q3F + S163P + S186R + Q215K + S225G + N227T + N252P + N267T +F276A + 80.36 79.76 78.52 S279G + K308E + K309E + K312E H15Y + A49W +N59E + S163P + S186R + S225G + N227T + E232D + N252P + 80.42 81.57 78.94N260Q + H272P + S279D + Y281P + S288P + K308Q + K309E + K312Q A49W +N59E + S163P + S186R + S225G + N227T + E232D + N252P + H272V + 80.7680.61 79.42 F276A + S279G + K308E + K309E + K312E H15Y + A49W + N59E +S163P + S186R + S225G + N227T + E232D + G235W + 80.94 81.12 79.29N252P + N260Q + H272V + S279D + Y281P + K308Q + K309E + K312Q S163P +S186R + Q215K + N227T + E232D + N252P + 80.97 80.09 79.23 N267T +F276A + S279G + Y281P + K308E + K309E + K312E Q3F + A49W + N59E +S163P + S186R + N227T + E232D + N252P + F276A + 80.99 80.56 79.67S279G + Y281P + K308E + K309E + K312E H15Y + A49W + N59E + S163P +S186R + S225G + N227T + E232D + N252P + 81.15 81.32 79.56 N260Q +H272P + S279D + Y281P + K308Q + K309E + K312Q A49W + N59E + S163P +S186R + S225G + N227T + E232D + N252P + F276A + 81.16 81.06 79.84S279G + Y281P + K308E + K309E + K312E H15Y + A49W + N59E + S163P +S186R + S225G + N227T + E232D + N252P + 81.17 80.92 77.86 N260Q +H272V + Y281P + K308Q + K309E + K312Q Q3F + T17W + A49W + N59E + S163P +S186R + D207N + T218Q + S225G + 81.18 80.71 78.60 N227T + G235W +N252P + H272P + Y281P + K308Q + K309E + K312Q H15Y + T17W + A49W +N59E + S163P + S186R + S225G + N227T + E232D + 81.24 81.19 77.87 G235W +N252P + N260Q + H272V + S279D + Y281P + K308Q + K309E + K312Q Q3F +A49W + N59E + S163P + S186R + S225G + N227T + N252P + F276A + 81.7681.55 80.60 S279G + Y281P + K308E + K309E + K312E Q3F + A49W + N59E +S163P + S186R + S225G + N227T + E232D + N252P + 81.81 81.59 80.51H272V + F276A + S279G + K308E + K309E + K312E A49W + N59E + S163P +S186R + N227T + E232D + N252P + 81.87 81.46 80.62 H272V + F276A +S279G + Y281P + K308E + K309E + K312E S163P + S186R + S225G + N227T +E232D + N252P + 81.93 83.27 81.14 N267T + F276A + S279G + Y281P +K308E + K309E + K312E H15Y + T17W + A49W + N59E + S163P + S186R +S225G + N227T + E232D + 81.96 81.81 79.45 N252P + N260Q + H272V +S279D + Y281P + K308Q + K309E + K312Q Q3F + S163P + S186R + Q215K +N227T + E232D + N252P + 82.12 81.36 80.19 N267T + H272V + F276A +S279G + K308E + K309E + K312E Q3I + H15Y + A49W + N59E + S163P + S186R +S225G + 82.13 81.51 79.96 N227T + E232D + G235W + N252P + N260Q +H272V + S279D + Y281P + K308Q + K309E + K312Q S163P + S186R + Q215K +S225G + N227T + E232D + 82.15 81.36 80.60 N252P + N267T + F276A +S279G + Y281P + K308E + K309E + K312E S163P + Q215K + S225G + N227T +E232D + N252P + 82.16 81.33 80.43 N267T + F276A + S279G + Y281P +K308E + K309E + K312E S163P + S186R + Q215K + S225G + N227T + E232D +82.28 81.08 79.79 N252P + N267T + H272V + F276A + S279G + K308E +K309E + K312E S163P + Q215K + S225G + N227T + E232D + N252P + 82.3981.76 80.60 N267T + H272V + F276A + S279G + K308E + K309E + K312E A49W +N59E + S163P + S186R + S225G + N227T + N252P + H272V + 82.60 82.50 81.14F276A + S279G + Y281P + K308E + K309E + K312E Q3F + S163P + S186R +Q215K + N227T + E232D + N252P + N267T + 82.61 83.55 81.68 F276A +S279G + Y281P + K308E + K309E + K312E Q3F + S163P + Q215K + N227T +E232D + N252P + N267T + F276A + 82.69 81.60 80.72 S279G + Y281P +K308E + K309E + K312E Q3F + H15Y + T17W + A49W + N59E + S163P + S186R +82.78 83.35 80.07 T218Q + S225G + N227T + G235W + S237W + N252P +H272P + Y281P + K308Q + K309E + K312Q S163P + Q215K + N227T + E232D +N252P + N267T + 82.84 84.19 82.34 H272V + F276A + S279G + Y281P +K308E + K309E + K312E Q3F + S163P + Q215K + S225G + N227T + E232D +N252P + 82.91 81.69 80.47 N267T + H272V + F276A + S279G + K308E +K309E + K312E Q3F + A49W + N59E + S163P + S186R + N227T + E232D + 82.9482.48 81.61 N252P + H272V + F276A + S279G + Y281P + K308E + K309E +K312E Q3F + S163P + S186R + Q215K + S225G + N227T + N252P + N267T +83.18 82.41 81.62 F276A + S279G + Y281P + K308E + K309E + K312E Q3F +H15Y + T17W + A49W + N59E + S163P + S186R + 83.42 84.07 80.61 D207N +T218Q + S225G + N227T + G235W + S237W + N252P + H272P + Y281P + K308Q +K309E + K312Q S163P + S186R + Q215K + S225G + N227T + N252P + 83.4784.70 83.22 N267T + H272V + F276A + S279G + Y281P + K308E + K309E +K312E S163P + Q215K + S225G + N227T + N252P + N267T + 83.50 82.53 81.62H272V + F276A + S279G + Y281P + K308E + K309E + K312E Q3F + S163P +Q215K + N227T + E232D + N252P + N267T + H272V + 83.62 82.52 82.27F276A + S279G + Y281P + K308E + K309E + K312E Q3F + A49W + N59E +S163P + S186R + S225G + N227T + E232D + 83.65 82.73 81.76 N252P +F276A + S279G + Y281P + K308E + K309E + K312E Q3F + A49W + N59E +S163P + S186R + S225G + N227T + N252P + H272V + 83.85 83.63 82.18F276A + S279G + Y281P + K308E + K309E + K312E A49W + N59E + S163P +S186R + S225G + N227T + E232D + N252P + H272V + 83.97 83.44 82.73F276A + S279G + Y281P + K308E + K309E + K312E S163P + Q215K + S225G +N227T + E232D + N252P + 84.12 82.91 82.08 N267T + H272V + F276A +S279G + Y281P + K308E + K309E + K312E Q3F + S163P + S186R + Q215K +S225G + N227T + E232D + N252P + N260Q + 84.21 84.61 82.56 N267T +H272P + S279D + Y281P + K308Q + K309E + K312Q Q3F + S163P + S186R +Q215K + N227T + E232D + N252P + N267T + H272V + 84.33 83.00 82.34F276A + S279G + Y281P + K308E + K309E + K312E S163P + S186R + Q215K +S225G + N227T + E232D + 84.55 85.90 84.65 N252P + N267T + H272V +F276A + S279G + Y281P + K308E + K309E + K312E Q3F + S163P + S186R +Q215K + S225G + N227T + N252P + N267T + H272V + 84.62 83.59 82.73F276A + S279G + Y281P + K308E + K309E + K312E Q3F + H15Y + V140I +S163P + S186R + Q215K + S225G + N227T + E232D + 84.79 83.58 82.43G235W + N252P + N260Q + N267T + H272V + S279D + Y281P + K308Q + K309E +K312Q Q3F + T17W + S163P + S186R + Q215K + S225G + N227T + E232D +G235W + 84.80 84.93 83.17 N252P + N267T + H272V + F276A + S279G +Y281P + K308E + K309E + K312Q Q3F + S163P + S186R + Q215K + S225G +N227T + E232D + N252P + N267T + 85.08 85.62 83.87 H272V + F276A +S279G + Y281P + K308E + K309E + K312Q Q3F + A49W + N59E + S163P +S186R + S225G + N227T + E232D + N252P + 85.15 84.60 83.54 H272V +F276A + S279G + Y281P + K308E + K309E + K312E Q3F + S163P + S186R +Q215K + S225G + N227T + E232D + N252P + N267T + 85.19 84.99 83.88H272V + F276A + S279G + Y281P + K308E + K309E + K312E Q3F + H15Y +T17W + S163P + S186R + Q215K + S225G + N227T + E232D + 85.20 85.06 83.15G235W + N252P + N260Q + N267T + H272V + S279D + Y281P + K308Q + K309E +K312Q Q3F + S163P + Q215K + S225G + N227T + E232D + N252P + N267T +H272V + 85.34 86.79 84.79 F276A + S279G + Y281P + K308E + K309E + K312EQ3F + H15Y + S163P + S186R + Q215K + S225G + N227T + E232D + G235W +85.53 85.89 84.59 N252P + N260Q + N267T + H272P + S279D + Y281P +K308Q + K309E + K312Q Q3F + A49W + N59E + S163P + S186R + S225G +N227T + E232D + G235W + 85.71 87.50 86.35 N252P + N260Q + H272P +F276A + S279D + Y281P + K308E + K309E + K312Q Q3F + H15Y + T17W +S163P + S186R + Q215K + S225G + N227T + E232D + 86.24 86.09 83.78N252P + N260Q + N267T + H272V + S279D + Y281P + K308Q + K309E + K312QQ3F + H15Y + S163P + S186R + Q215K + S225G + N227T + E232D + N252P +86.42 87.01 85.52 N260Q + N267T + H272P + S279D + Y281P + K308Q +K309E + K312Q Q3F + H15Y + S163P + S186R + Q215K + S225G + N227T +E232D + G235W + 86.61 86.96 85.35 N252P + N267T + H272V + F276A +S279G + Y281P + K308Q + K309E + K312Q Q3F + H15Y + T17W + S163P +S186R + Q215K + S225G + N227T + E232D + 86.96 86.83 84.64 N252P +N267T + H272V + F276A + S279G + Y281P + K308Q + K309E + K312Q Q3F +H15Y + A49W + N59E + V140I + S163P + S186R + Q215K + S225G + 87.16 85.9284.43 N227T + E232D + G235W + N252P + N260Q + N267T + H272V + S279D +Y281P + K308Q + K309E + K312Q

Table 8+8A show that adding substitutions mentioned in this invention toSEQ ID NO: 1 generally increase the Tm (melting temperature) value bothin a buffer system and in buffer supplemented with the detergent ModelA. A Tm increase is here used as evaluation for increased stability ofthe variants.

TABLE 9 Thermostability for variants comprising di-sulphide bridgesEffect of A269C − Y282C nDSF Delta Delta Di- Tm (° C.) Tm (° C.) Tm Tmsulphide 0 g/L 2 g/L 0 g/L 2 g/L Substitution Base Substitution BridgeModel A ModelA Model A Model A A49W + F276A + A49W + F276A + No 58.7958.14 5.30 5.59 K308E K308E Disulfide Bridge A49W + A269C + A49W +F276A + A269C + 64.09 63.73 F276A + Y282C + K308E Y282C K308E A49W +N227T + A49W + N227T + No 58.97 56.27 5.73 7.01 F276A F276A DisulfideBridge A49W + N227T + A49W + N227T + A269C + 64.70 63.28 A269C + F276A +F276A Y282C Y282C A49W + N227T + A49W + N227T + No 63.06 63.21 5.66 4.83F276A + K308E F276A + K308E Disulfide Bridge A49W + N227T + A49W +N227T + A269C + 68.72 68.04 A269C + F276A + F276A + K308E Y282C Y282C +K308E A49W + N227T + A49W + N227T + A269C + 64.5 63.08 ND ND A269C +Y282C + K308E Y282C K308E A49W + S186R + A49W + S186R + No 60.13 56.724.32 6.39 N227T + F276A N227T + F276A Disulfide Bridge A49W + S186R +A49W + S186R + A269C + 64.45 63.11 N227T + A269C + N227T + F276A Y282CF276A + Y282C A49W + S186R + A49W + S186R + No 63.55 62.46 5.20 6.18N227T + F276A + N227T + F276A + Disulfide K308E K308E Bridge A49W +S186R + A49W + S186R + A269C + 68.75 68.64 N227T + A269C + N227T +F276A + Y282C F276A + Y282C + K308E K308E A49W + S186R + A49W + S186R +No 59.72 54.84 4.88 9.24 N227T + K308E N227T + K308E Disulfide BridgeA49W + S186R + A49W + S186R + A269C + 64.60 64.08 N227T + A269C +N227T + K308E Y282C Y282C + K308E A49W + Y124R + A49W + Y124R + No 63.3062.69 5.25 5.54 N227T + F276A + N227T + F276A + Disulfide K308E K308EBridge A49W + Y124R + A49W + Y124R + A269C + 68.55 68.23 N227T + A269C +N227T + F276A + Y282C F276A + Y282C + K308E K308E A49W + Y124R + A49W +Y124R + No 61.89 57.85 2.76 6.66 S186R + N227T + S186R + N227T +Disulfide F276A F276A Bridge A49W + Y124R + A49W + Y124R + A269C + 64.6564.51 S186R + N227T + S186R + N227T + Y282C A269C + F276A + F276A Y282CA49W + Y124R + A49W + Y124R + No 64.61 62.60 4.17 5.86 S186R + N227T +S186R + N227T + Disulfide F276A + K308E F276A + K308E Bridge A49W +Y124R + A49W + Y124R + A269C + 68.78 68.46 S186R + N227T + S186R +N227T + Y282C A269C + F276A + F276A + K308E Y282C + K308E A49W + Y124R +A49W + Y124R + No 61.72 56.68 4.02 7.91 S186R + N227T + S186R + N227T +Disulfide K308E K308E Bridge A49W + Y124R + A49W + Y124R + A269C + 65.7464.59 S186R + N227T + S186R + N227T + Y282C A269C + Y282C + K308E K308EN227T + F276A + N227T + F276A + No 63.38 62.25 4.86 5.91 K308E K308EDisulfide Bridge N227T + A269C + N227T + F276A + A269C + 68.24 68.16F276A + Y282C + K308E Y282C K308E S186R + F276A + S186R + F276A + No59.16 58.33 4.88 4.81 K308E K308E Disulfide Bridge S186R + A269C +S186R + F276A + A269C + 64.04 63.14 F276A + Y282C + K308E Y282C K308ES186R + N227T + S186R + N227T + No 59.58 57.18 5.17 6.47 F276A F276ADisulfide Bridge S186R + N227T + S186R + N227T + A269C + 64.75 63.65A269C + F276A + F276A Y282C Y282C S186R + N227T + S186R + N227T +A269C + 68.74 68.43 ND ND A269C + F276A + F276A + K308E Y282C Y282C +K308E S186R + N227T + S186R + N227T + No 58.87 54.49 5.55 8.88 K308EK308E Disulfide Bridge S186R + N227T + S186R + N227T + A269C + 64.4263.37 A269C + Y282C + K308E Y282C K308E Y124R + S186R + Y124R + S186R +No 63.60 62.60 4.88 5.71 N227T + F276A + N227T + F276A + Disulfide K308EK308E Bridge Y124R + S186R + Y124R + S186R + A269C + 68.48 68.31 N227T +A269C + N227T + F276A + Y282C F276A + Y282C + K308E K308E

Table 9 shows, that addition of the disulfide bride A269C+Y282 increasesthe Tm (melting temperature) values both in a buffer system and inbuffer supplemented with the detergent Model A (approx. 2-10° C.=DeltaTm) for all the tested variants (Base mutation +/−Disulfide bridge). ATm increase are here used as evaluation for increased stability of thevariants.

1. A dispersin variant, comprising an alteration at one or moreposition(s) corresponding to position(s): 2, 3, 12, 15, 17, 18, 19, 22,23, 24, 25, 26, 30, 32, 34, 43, 44, 45, 49, 52, 54, 56, 57, 58, 59, 60,62, 63, 67, 68, 71, 72, 74, 77, 79, 80, 81, 82, 90, 99, 100, 103, 104,105, 106, 107, 110, 111, 113, 114, 116, 117, 118, 119, 120, 122, 123,124, 125, 126, 127, 128, 131, 135, 138, 139, 140, 142, 145, 147, 148,149, 150, 151, 152, 163, 164, 167, 168, 170, 171, 173, 174, 175, 177,178, 179, 181, 185, 186, 187, 188, 189, 199, 200, 203, 204, 205, 207,208, 210, 215, 217, 218, 221, 222, 224, 225, 227, 230, 232, 233, 234,235, 237, 244, 249, 251, 252, 253, 254, 256, 260, 261, 262, 263, 264,265, 267, 268, 270, 271, 272, 273, 274, 276, 278, 279, 280, 281, 282,283, 284, 287, 288, 290, 291, 296, 300, 301, 303, 304, 305, 306, 308,309, 312, 314, 315, 319, 321 or 323 of the polypeptide shown in SEQ IDNO: 1, wherein the dispersin variant has hexosaminidase activity andwherein the variant has increased thermo stability measured as half-lifeimprovement factor, HIF, compared to the dispersin with SEQ ID NO:
 1. 2.A dispersin variant, comprising at least one alteration selected fromthe group consisting of: D2A, D2L, D2N, D2R, D2V, D2W, Q3F, Q3I, Q3L,Q3M, Q3P, Q3V, Q3Y, Q3T, S12A, H15F, H15Y, T17W, T17C, T17E, T17F, T17M,T17R, T17V, V18L, E19D, E19N, E19P, K22A, K22M, K22V, S23C, S23E, S23I,S23L, S23R, S23T, S23V, V25R, D26M, Y30*, Y30D, Y30L, Y30M, Y30N, Y30R,Y30T, Y30V, G32L, G32M, G32R, N43*, N43H, N43L, E44*, N45D, N45L, N45V,A49W, A49Y, Y52*, Y52M, G54L, G54M, G54N, S56T, S56W, S57W, E58N, N59A,N59C, N59D, N59E, N59F, N59M, N59R, N59V, N59W, T60V, N62C, N62D, N62H,N62Q, N62W, T63C, T63D, T63L, T63N, T63R, T63V, K67A, K67L, N68L, N68Q,L71H, L71N, L71R, L71V, L71W, S72*, S72C, S72D, S72E, S72F, S72G, S72I,S72M, S72N, S72R, S72T, S72Y, I74L, S77A, D79V, K80*, K80E, K80H, K80L,K80N, K80Q, K80V, K80W, D81A, D81G, D81L, D81R, D81S, D81T, D81V, D81W,I82V, L90F, E99Q, E99R, L100S, K103A, K103R, K104N, K104W, D105N, V106A,V106D, V106E, V106H, V106K, V106L, V106M, V106N, V106Q, V106W, V106Y,K107A, K107C, K107L, K107M, K107T, K107V, K107W, N110M, N110R, N110V,D111A, D111E, D111M, D111N, D111Q, D111R, D111V, D111W, V113T, T114C,T114S, Y116D, Y116N, Y116R, S117*, S117D, S117H, S117N, S117P, E118*,E118A, E118D, E118G, E118L, E119G, E119W, T120I, T120L, T120M, T120V,T120W, D122*, D122H, D122R, Y123W, Y124C, Y124I, Y124K, Y124L, Y124M,Y124Q, Y124R, Y124T, Y124V, Y124W, D125C, D125G, D125K, D125Q, D125R,N126V, R127D, R127H, R127K, R127L, R127M, R127Q, R127W, V128C, V128L,V128T, D131V, Q135*, Q135A, Q135D, Q135E, Q135K, Q135M, Q135Y, D138K,D138L, D138M, D138Q, D138R, D138S, D138V, D138W, E139W, D142R, D142W,Y145*, Y145H, Y145L, Y145N, Y145V, P147A, P147C, P147D, P147F, P147G,P147L, P147M, P147R, P147S, P147T, P147V, K148A, K148D, K148L, K148V,F149L, F149M, F149N, E150D, E150H, E150K, E150L, E150M, E150N, E150R,E150V, E150W, E150Y, G151A, G151C, G151D, G151L, G151N, G151P, G151S,G151W, K152D, K152L, K152R, G164D, G164E, G164H, G164S, G164V, V167D,V167E, V167L, V167P, V167Q, V167R, V167W, H168N, L170A, L170D, L170E,L170F, L170H, L170K, L170M, L170N, L170P, L170Q, L170R, L170S, L170V,L170W, L170Y, D171A, D171C, D171E, D171K, D171L, D171M, D171Q, D171R,D171V, D171W, D171Y, I173C, D174H, D174M, D174N, D174V, D174W, F175Y,N177M, Q178*, Q178A, Q178K, Q178R, Q178W, I179T, S181C, S181D, S181F,S181G, S181N, S181P, S181Q, S181T, S181V, S181W, E185M, E185R, E185V,E185W, S186D, S186E, S186H, S186I, S186K, S186L, S186M, S186N, S186Q,S186R, S186V, S186W, K187C, K187D, K187G, K187R, K187S, K187V, K187W,Y188P, E189L, E189V, E189W, S199C, S199L, S199M, S199Y, E200D, E200F,E200K, E200L, E200M, E200N, E200R, E200W, A203C, A203D, A203E, A203G,A203L, A203M, A203P, A203R, A203S, A203T, A203V, A203W, N204L, N204M,N204V, N204W, N204Y, L205I, D207A, D207C, D207E, D207G, D207N, D207Q,D207S, D207V, D207W, S208A, S208C, S208D, S208G, S208L, S208Q, S208T,S208V, S208W, S210T, Q215R, Q215M, Q215L, Q215*, S217V, T218A, T218L,T218Q, T218R, T218V, G222D, E224A, E224P, N227A, N227Q, N227R, N227S,N227T, N227K, D230*, D230R, D230T, D230W, E232D, E232V, N233H, N233Q,N233R, N233W, W234R, G235W, G235A, G235E, G235F, G235H, G235I, G235L,G235M, G235N, G235P, G235S, G235V, S237C, S237G, S237M, S237N, S237W,S237Y, Y244C, Y244E, Y244M, L249H, L249K, L249Q, L249R, L249W, L249Y,S251L, S251N, S251R, S251W, N252P, N252C, G253D, G253W, F254I, F254L,F254M, F254Y, Q256E, Q256R, N260*, N260A, N260C, N260E, N260I, N260K,N260L, N260M, N260Q, N260R, N260V, N260W, N260Y, E261*, E261A, E261D,E261R, E261W, Q262*, Q262F, Q262H, Q262W, Q262Y, M263K, M263L, M263Q,D264*, D264C, D264E, Y265F, N267S, N267T, W268C, W268E, W268M, W268R,Y270F, A271D, A271G, H272D, H272I, H272M, H272P, H272V, H272W, N273W,K274R, K274A, K274H, F276A, F276C, F276K, F276N, F276G, F276L, F276M,F276P, F276S, F276V, F276W, I278A, I278K, I278N, I278Q, I278V, S279C,S279D, S279E, S279G, S279N, D280C, D280E, Y281*, Y281A, Y281C, Y281H,Y281K, Y281N, Y281P, Y281R, Y282E, Y282N, H283I, A284I, A284L, A284N,A284P, A284T, A284V, T287N, S288D, S288K, S288N, V290I, K291L, K291R,K291V, T296C, E300A, E300D, H301C, H301N, H301R, T303A, T303C, T303G,T303K, T303Q, T303R, T303W, D304C, D304M, L305M, L305N, S306C, K308A,K308D, K308G, K308I, K308L, K308Q, K308S, K308T, K308V, K308Y, K309A,K309C, K309D, K309H, K309L, K309M, K309N, K309Q, K309S, K309T, K309I,K312A, K312L, K312M, K312N, K312Q, K312S, K312W, E314I, E314L, E314V,L315I, L315V, R319A, and N323R, wherein the positions correspond to thepositions of the polypeptide shown in SEQ ID NO: 1, wherein thedispersin variant has hexosaminidase activity preferably beta-1,6N-acetylglucosaminidase activity.
 3. The dispersin variant according toclaim 2, wherein the alteration at the one or more position(s) producesa dispersin variant having increased thermo-stability measured asimprovement factor, HIF, of at least 1.1 compared to the polypeptideshown in SEQ ID NO:
 1. 4. The dispersin variant according to claim 1,wherein the variant has at least 60% sequence identity to thepolypeptide shown in SEQ ID NO:
 1. 5. The dispersin variant according toclaim 1, wherein the variant is obtained from Terribacillus.
 6. Thedispersin variant according to claim 1, wherein the variant comprisesone or more motifs: GXDE (SEQ ID NO 8),[EQ][NRSHA][YVFL][AGSTC][IVLF][EAQYN][SN] (SEQ ID NO9),[VIM][LIV]G[GAV]DE[VI][PSA] SEQ ID NO 10), [WND[SQR][IVL][TLVM] (SEQID NO 11), QSTL (SEQ ID NO 12), NKFFY (SEQ ID NO 13) or NLD[DR]S (SEQ IDNO 14).
 7. The dispersin variant according to claim 1, wherein thedispersin variant has improved stability, measured as having animprovement factor e.g. RAR or HIF>1.0, compared to the dispersin havingthe polypeptide sequence shown in SEQ ID NO:
 1. 8-16. (canceled)
 17. Acomposition, comprising a variant according to claim
 1. 18. A method ofcleaning an item comprising washing the item with the composition ofclaim 17, wherein the item is a textile or a hard surface. 19-23.(canceled)